| Program and abstracts | Poster presentations |
|---|
Pathological morphology of the human and animal diseases
Iatrogenic, environmental and experimental pathology
“Herman Jurak” round table on rheumatological diseases
Ethical forum – Forensic pathology:
Dental pathology
Short courses in cytopathology
IN VITRO DIFFERENTIATION AND NEURAL TRANSPLANTATION OF EMBRYONIC STEM CELLS: NEW STRATEGIES FOR GENE TRANSFER INTO THE NERVOUS SYSTEM
S. Brandner, Y. Benninger, A. Cozzio, S. Marino, A. Aguzzi
Institute of Neuropathology, University Hospital, Zurich, Switzerland
Several strategies can be employed to study the function of specific genes in the central nervous system. Although the generation of transgenic and knockout mice is one of the most effective approaches, it may prove laborious and expensive, in particular if numerous mutations are to be assessed. We therefore employed the neural transplantation strategy, which provides more flexibility than the transgenic approach and proved valuable in the investigation of neurodegeneration 1, 2. However, instead of using committed neural precursor cells (which are derived from midgestation embryos and are usually scantily permissive to any gene transfer), we used embryonic stem (ES) cells, which are omnipotent and allow for efficient in vitro gene transfer by electroporation. Following successful integration of desired constructs, these cells can be clonally selected and amplified to large quantities. To attain neural grafts, ES cells were first cultured under specifically adapted conditions in vitro, leading to the formation of aggregates (“neurospheres”). Following several days in vitro culture, such neuronally differentiated spheres were then stereotaxically transplanted into the brain of recipient adult mice, where they form neurally differentiated tissue. The flexibility of this approach will allow us to study the dynamics of neurodegenerative diseases and also the impact of gene disruptions on neural development.
References
- Brandner, S. et al. Normal host prion protein necessary for scrapie-induced neurotoxicity. Nature 379, 339-43 (1996).
- Brandner, S. et al. Normal host prion protein (PrPC) is required for scrapie spread within the central nervous system. Proc Natl Acad Sci U S A 93, 13148-51 (1996).
TUBERCULOUS MENINGOENCEFALITIS: [RE]EMERGING PATHOLOGY
R Bussani, P. Cusati, N. Pavletić, F. Silvestri
Institute of Pathological Anatomy, University of Trieste, Italy
Since 1940, the incidence of tuberculous meningoencefalitis has declined markedly in many western European countries and in USA. However, during the last years, an increasing proportion of tuberculosis [TB] notifications is reported, but very few studies are based on autopsy material. In Trieste, where the autopsy rate is presently almost 80% of all hospital deaths, we have a very large volume of reports of autopsies. We are, therefore, able to study changes in some of the main human pathological conditions retrospectively between 1901 till today.
In 1901, tuberculosis caused death of 22.4% of subjects and 41.6% of people who died before the age of 40. A tubercular involvement of the central nervous system was evidenced at the autopsy of almost 37% of the affected patients with hematogenous dissemination. The introduction of chest X-ray examinations, improved living conditions and streptomycin and isoniazid introduction resulted with a significant reduction of death rates from TB: in 1993 the autopsy rate of TB was 0,66%.
In the last five years there has been a significant, progressive increase of active TB in autopsy frequency.
In 1998, 22 out of 1740 autopsies of adult patients (1.26%) were found to have a pulmonary or systemic TB (mostly miliaries or tubercular bronchopneumonia) and in almost 80% of these cases the lesion was the underlying cause of death. Tuberculous meningitis or a specific meningoencefalitis was evidenced in 32% of the affected subjects. In these patients clinical signs and symptoms often weren’t very clear and didn’t reach the “minimum standard” necessary to request the microbiologic isolation of the Micobacterium Tuherculosis. There was no clinical diagnosis or clinical suspect in any of these patients.
These facts emphasise the need for improved surveillance of TB in Trieste as well as in other parts of the developed world.
3
SUBTOTAL AGENESIA OF SPINAL ROOTS IN A TWIN: A VERY RARE MALFORMATION PROBABLY DUE TO A FETAL INFECTION
R. Bussani, M. Pellegrino, N. Pavletic, F. Silvestri
Institute of Pathological Anatomy, University of Trieste, Italy
Infections of the fetus may cause two types of lesions. Firstly, inflammatory and destructive changes caused by the infectious process may occur. Secondly, the tissue damage resulting from the infection may derange subsequent development, leaving malformations in its wake. These components may coexist or overlap.
We report a very rare malformation of the spinal cord and spinal roots in one of two twins (dichorionic diamniotic) spontaneously aborted at 20 weeks’ gestation by a 33-ycar-old grava 2, para 1 mother who developed a “flow-like syndrome” during the first trimester.
Both of the fetuses were females and weighed about 500 grams.
Macroscopically the placentae presented focal accentuated granularity of the villous tissue with a laminar thickening of the maternal zone of the disks. The microscopic features were not pathognomonic of any specific infection, the main features being a relative hypoplasia with dysmaturity of villous stems, increased perivillous fibrin, and an acute, necrotising, and multifocal chorioamnianitis.
At post-mortem examination one fetus was absolutely normal. The other presented a moderate cerebral ventriculomegalia and, during the extraction of the brain from the cranial cavity, the spinal cord spontaneously came out of the spinal channel. The spinal cord was lacking in nervous spino-gangliar connections at the cervical and dorsal level.
To our knowledge, this is the first morphohistological description of a congenital anomaly of the spinal cord of this type.
4
BRAIN PATHOLOGY OF SLOVENIAN AIDS PATIENTS
M. Volavšek, M. Popović
Institute of Pathology, Medical Faculty, Ljubljana, Slovenia
Introduction
The central nervous system (CNS) is, besides the lymphoid system, the major target of HIV infection. The only cell-type containing HIV in CNS is microglia which becomes infected via macrophages from the peripheral blood. In addition to opportunistic infections, neoplasms, and encephalitis/encephalopathy directly caused by HIV, several changes, indirectly caused by HIV, can occur. These include diffuse poliodystrophy, myelin pallor, aseptic meningitis, vacuolar myelopathy, and peripheral neuropathies. Neuronal damage is believed to be caused by cytotoxic factors released by infected microglia.
Aim of the study. The aim of our retrospective study was to disclose which neuropathologic changes dominate in Slovenian AIDS patients. We examined brains of all AIDS patients autopsied at the Institute of Pathology, Medical Faculty in Ljubljana from 1989-99.
Methods
Neuropathologic examination has been performed in 20 cases using standard protocol, and H&E and Klüver-Barrera staining. Histohemical and immunohistochemical methods, as well as in situ hybridisation have been performed to detect HIV and eventual opportunistic microorganisms.
Results
Only one of 20 patients showed no brain pathology. Opportunistic infections were present in 12 patients (toxoplasmosis 4/12, cryptococcosis 2/12, cytomegalovirus infection 3/12, morbili 1/12, progressive multifocal leukoencefalopathy 2/12). In 5 patients encephalopathy not otherwise specified has been diagnosed. One patient had primary CNS high-grade lymphoma (large B-cell lymphoma), and another one HIV encephalopathy.
Conclusion. The predominance of opportunistic infections in brain pathology of our AIDS patients is in concordance with reports from the literature. The consequences of direct and indirect effects of HIV have been present rarely. In the cases of encephalophathy not otherwise specified we were not able to identify eventual microorganisms. More sophisticated detection methods as polymerase chain reaction, at present not available for routine use at our institution, would probably enable us to classify these cases.
5
SURGICAL REMOVAL OF THE RIGHT CEREBRAL HEMISPHERE: AN ANATOMIC ANALYSIS OF THE BRAIN TWENTY YEARS LATER
R. Bussani, M. Pellegrino, F. Bassan, N. Pavletić, F. Silvestri
Institute of Pathological Anatomy, University of Trieste, Italy
The patient was a female suffering from hypoxic perinatal cerebropathy. At the age of 35 epileptic seizures began periodically. For some years the fits were partially relieved by pharmacological therapy, but as time passed epileptic events increased in frequency and length, and pharmacological therapy became useless.
At age 41 the patient went abroad where she underwent to a complex neurosurgical operation which included surgical removing of the right cerebral hemisphere. The post-surgery status showed decreased performance with a significant loss of motor and intellectual ability, but the patient was able to fulfil easy tasks. She died twenty years later of a poorly differentiated adenocarcinoma of the extrahepatic bile ducts metastatic to liver and abdominal lymph nodes and multiple liver abscesses.
Autopsy showed a conspicuous internal thickening of the frontal cranial bone (mm. 17). The brain weight was 530 grams. The residual cerebral hemisphere appeared generally atrophied. The histopathological analysis evidenced a severe neuronal loss in layers of the cerebral cortex and of the brain nuclei with diffuse reticular formation. The cerebellar cortex showed a decrease in number of Purkinje cells and ballooning of most of the remaining cells. A detailed photographic report of the brain was performed.
To our knowledge, this is the first macroscopically documented case of the brain that underwent such a destructive type of surgery so many years before death.
6
ORGANIZATION OF CORTICAL CIRCUITRY IN A CASE OF SEMILOBAR HOLOPROSENCEPHALY; RAPID GOLGI STUDY
M. R. Rašin, Z. Petanjek*, D. Jukić, K. Kostović, S. Jukić, B. Krušlin, I. Kostović
*Croatian Institute for Brain Research, Zagreb, Croatia
School of Medicine, Zagreb, Croatia
Holoprosencephaly is a cerebral malformation characterised by inadequate division of the prosencephalon. Gross morphological abnormalities in a case of semilobar holoprosencephaly (36-week-old fetus) suggest that disturbance in neural development occurs between the 6th and 8th embryonic week. At this time the first cell containing nonproliferative layer (preplate layer) becomes divided by migrating neurones which build the cortical plate, into the upper located marginal zone, and below the cortical plate located subplate zone. Our hypothesis was that histological slices would show disturbances in normal development of transitory fetal cortical layers (subplate) and disarrangement in circuitry organisation between neurones of cortical plate. Using rapid Golgi methods we have tried to answer following questions: 1. Does the preplate divide into marginal zone and primordial subplate? 2. Could the subplate be detected below the cortical plate? 3. Is there an inversion of layers in cortical plate? Numerous subplate like neurones in marginal zone suggest that there was no clear dividing of preplate, and that the majority of preplate neurons is located in the marginal zone of a holoprosencephalic brain. This finding suggests that semilobar holoprosencephaly could be the human reeler mouse like model. Surprisingly, the consequences of abnormal preplate development present in reeler mouse (absence of subplate and inversion of cortical layers) were not present, and the organisation of cortical circuitry in cortical plate and subplate layer was amazingly normally developed according to the huge malformations of gross morphology. This investigation shows that malformation of brain development could be a model for understanding histogenetic events during normal development, especially in the context of human developmental specialities, which cannot be understood by using animal experimental models.
7
REPORT ON RARE TUMORS AND TUMOR-LIKE CONDITIONS OF THE PITUITARY (WITH A 10-YEAR REVIEW)
J. Talan-Hranilović, M. Lechpammer, Ž. Gnjidić, M. Kalousek
Department of Clinical Pathology “Ljudevit Jurak”,Clinic of Neurosurgery and Clinic of Radiology University Hospital “Sestre milosrdnice”, Zagreb, Croatia
The objective of this work was to analyse 10 years of experience of the Clinical Department of Pathology “Ljudevit Jurak” in pituitary expansive neoplastic processes diagnostics. Our special attention was paid to rare pituitary tumours. Various types of pituitary tumours were evaluated according to age and sex, with exception of pituitary adenoma.
Study was designed as a retrospective review of the records of patients who underwent surgery of pituitary tumours between January 1989 and February 1999. The distribution of tumours was analysed according to age at surgery, sex, size and patohistological type of the tumour.
Total of 238 pituitary tumours was diagnosed during the last 10 years. The most common conditions were Adenoma glandulae hypophiseos (95,8%). Rare conditions were diagnosed in 10 cases (4,2%).
Average age at diagnosis was 43,4 years. Male to female ratio was 1:1 . Other rare conditions of the pituitary are analysed in the Table.
RARE CONDITIONS OF THE PITUITARY |
NUMBER OF CASES |
| 1. Inflammatio chronica | 4 (1,68%) |
| 2. Inflammatio chronica et microhyperplasia nodularis cellularum acidophilicum. | 1 (0,42%) |
| 3. Inflammatio suppurativa subacuta | 1 (0,42%) |
| 5. Adenohypophisitis lymphocytaria | 1 (0,42%) |
| 6. Gangliocytoma | 1 (0,42%) |
| 7. Paraganglioma | 1 (0,42%) |
| 8. Cystis epidermalis | 1 (0,42%) |
| TOTAL | 10 (4,2%) |
Average age of patients with rare pituitary tumours was 45,52 years, with male to female ratio of 1:1,3. In comparison with more common pituitary expansive processes it can be concluded that rare conditions emerge at any age and regardless to sex .
Until now only few studies have attempted such an analysis as the one presented here. Most of the published data suggested that frequency of different types of pituitary tumours vary according to age and sex. Our report confirms those observations with addition of possibility of diagnosing rare and unexpected conditions and tumours among any age group in both genders.
References:
- Bilbao JM, Horvath E, Kovach K et al. Intrasellar paraganglioma associated with hypopituitarism. Arch of Pathology and Lab Medicine 1978; 102(95):302
- Mindermann T, Wilson CB et al. Age-related and gender related occurrence of pituitary adenomas. Clin Endocrinol 1994 Sep;41(3):359
- Hashimoto M, Yanaki T, Nakahara N, Masuzawa T. Lymphocytic adenohypophisitis:an immunohistochemical study. Surg Neurol 1991 Aug;36(2):137
8
LHERMITTE-DUCLOS DISEASE: A CASE REPORT
L. Negovetić, V Beroš, V Čerina, T Sajko, J. Talan-Hranilović*.
Department of Neurosurgery , *“Ljudevit Jurak” Clinical Department of Pathology University Hospital “Sestre Milosrdnice”, Zagreb, Croatia
Lhermitte-Duclos disease or dysplastic cerebellar gangliocytoma is a rare lesion involving the cerebellum, characterised by enlarged cerebellar folia containing abnormal ganglion cells.
This case report describes a 24-year-old girl with signs of aqueductal stenosis. She presented to her local hospital with a 3-month history of occipital headache that became more severe and was associated with nausea and vomiting 2 weeks prior to admission.
CT scans showed only hypodense cerebellar lesion and MR imaging showed a left- sided cerebellar lesion extending to the brain steam with consequent aqueductal obliteration.
The cerebellar lesion consisted of abnormally thickened folia with abnormal hyperintensivity of the cerebellum on the T2- weighted images.
This lesion did not enhance with gadolinium.
The patient underwent a posterior fossa craniectomy and a CI laminectomy with total resection of the cerebellar lesion.
The postoperative course was normal and the patient showed improvement in her neurological status. Total removal of the tumour resolved the presenting symptoms completely.
Observation in patients with Lhermitte-Duclos disease is necessary because long term follow-up of this patients is essential to identify signs of Cowden’ s disease, which carries a risk of malignancy development.
Literature:
- Meltzer CC, Smirniotopoulos JG, Jones RV. The striated cerebellum: an MR imaging sign in Lhermitte-Duclos disease(dysplastic gangliocytoma). Radiology 1995;194;699-703.
- Siddiqi SN, Fehlings MG. Lhermitte-Duclos disease mimicking adult-onset aqueductal stenosis. J Neurosurg 1994;80;1095-1098.
- Poljakevic Z, Jadro-Šantel D, Bešenski N, Popović M, Pažanin L, Negovetić L. Dysplastic granulo-molecular hypertrophy of the cerebellar cortex(Lhermitte-Duclos disease): report of three cases. Neurologia 1990;39;259-271.
- Rimbou J, Isamat F. Dysplastic gangliocytoma of the cerebellum(Lhermitte-Duclos disease) and its relation to the multiple hamartoma syndrome(Cowden disease). J Neurooncol 1994;18;191-197.
9
DEMENTIAS – CLASIFFICATION, SYMPTOMS, DIAGNOSTICS AND TREATMENT POSSIBILITIES
V. Folnegović – Šmalc, V. Demarin*
Univ. Psych. Hospital Vrapče, Univ. Department of Neurology, Zagreb, Croatia
*“Sestre milosrdnice” Univ. Hospital, Zagreb, Croatia
There are various definitions of dementia. It is a disorder characterised both by memory and reasoning deterioration sufficient to impair personal activities of daily living. The memory impairment affects the registration, storage and retrieval of new information. However, all existing definitions have one main problem and that is lack of quantification.
Dementias are divided into early-onset (before age 65) and late-onset subtypes. Alzheimer’s disease (DAT) accounts for over one-half of all dementias. The most common subtype is caused by a vascular disease and is known as multi-infarct dementia (MID). Reversible dementias are those in which a specific trigger can be identified as responsible for intellectual decline and possibly be treated. Diagnosis is set apart from clinical symptoms and signs, by means of psychological testing, laboratory procedures and neuro-imaging modalities.
Treatment is still not satisfactory but new therapies are emerging.
10
CLASSIFICATION OF FELINE MALIGNANT LYMPHOMA – APPLICATION OF THE REAL CLASSIFICATION
K. Koehler*, A. Kipar**, M.L. Jackson**, M. Reinacher**
* Inst. Vet. Pathol. Univ. Giessen, Giessen, Germany
**. Dept. Vet. Pathol. Western College of Vet. Med. Univ. Saskatchewan, Sackatoon, Canada
Malignant lymphomas are frequent neoplasias in cats and are in a high percentage associated with feline leukaemia virus (FeLV) infection. Apart of the anatomical distribution, the histopathological characteristics, cellular origin can, also, be considered as features for feline malignant lymphoma classification.
Aim of the study
The “Revised European-American Classification of Lymphoid Neoplasms” (REAL classification), based on morphological features in connection with cellular origin, has been used for human malignant lymphomas classification. The purpose of this study was to apply the REAL classification to feline malignant lymphomas.
Methods
A total of 123 malignant lymphoma cases were immunohistologically examined for FeLV expression; 67 provided FeLV – positive, 56 FeLV – negative. Cases were classified according to the REAL classification, based on anatomical demonstration of B or T cell origin.
Results
Mediastinal lymphomas were mostly FeLV – positive and of T cell origin, whereas intestinal lymphomas were mostly FeLV – negative and of B cell origin. While major differences between FeLV – positive and – negative cases concerning the degree of cellular differentiation could not be stated, anatomical distribution and cellular origin were partly correlated with presence or lack of FeLV expression, respectively. Most cases could readily be sorted into the scheme of diagnosis, offered by the REAL classification.
Conclusion
Results establish the REAL classification as a useful scheme for feline malignant lymphoma classification.
11
VARIETY OF LESIONS IN FELINE INFECTIOUS PERITONITIS
A. Kipar*, W. Leukert**, S. Bellmann**, M. Reinacher*
*Institut für Veterinär-Pathologie, Universitet Giessen, Giessen, Germany
**Institut für Veterinär-Pathologie, Universitet Leipzig, Leipzig, Germany
Feline infectious peritonitis (FIP) is a widespread coronavirus (CoV)-induced disease in cats, which is characterised by variable combinations of fibrinous-granulomatous-necrotizing lesions with or without a serofibrinous serositis.
Aim of the study
The purpose of this study was to demonstrate the range of lesions, which develop after FIP virus (FIPV) infection in order to gain further information on the pathogenesis of FIP and to improve post-mortem diagnostic facilities.
Methods
Histopathology, immunohistology (demonstration of FIPV antigen and anti-FIPV antibodies in situ, characterisation of inflammatory cells), serology (demonstration of CoV antibodies and CoV-specific immune complexes), PCR and electron microscopy (demonstration of CoV particles) were applied and final diagnosis established at necropsy.
Results
FIP lesions comprised of the classical exudative and dry forms with fibrinous-granulomatous-necrotizing lesions in various organs, can also be restricted to leptomeningitis and chorioiditis extending into the brain and spinal cord, (chronic) granulomatous-necrotizing lymphadenitis of the mesenteric lymph nodes, and chronic pleuritis, characterised by minimal expression of viral antigens, B cell dominance, and the presence of multiple plasma cells positive for anti-FIPV antibodies in situ. However, systemic immune response is generally observed.
Conclusions
The study reveals FIP as a systemic disease, represented by generalised or local alterations. Diagnosis of FIP often requires immunohistological demonstration of CoV antigen in altered tissues in order to establish the diagnosis of FIP. Furthermore, results indicate that different types of immune reaction correspond with the different lesions observed.
12
NEUROPATHOLOGICAL SURVEY OF BOVINE BRAINS FROM ANIMALS WITH CLINICAL DIAGNOSIS OF NEUROLOGICAL SYMPTOMS
P. Juntes, M. Pogačnik
University of Ljubljana, Veterinary Faculty, Ljubljana, Slovenia
Aim
Since the beginning of bovine spongiform encephalopathy outbreak in 1986, interest for diseases of the central nervous system in cattle grew considerably, and many countries, including our, perform neuropathological surveys (1,2). The aim of this paper is to present pathological findings in brains of cattle with clinical history of neurological signs, examined in Slovenia since the year 1990.
Material and methods
Brains were examined in all adult cattle and calves reported to suffer neurological clinical signs. Routinely included gross pathology examination was performed, as well as pathohistology, bacteriological and virological examination. Brains were extracted from the cranium, smaller pieces of cerebrum, brain stem, cerebellum and medulla were taken for bacteriology and virology, and the rest was fixed in buffered formalin and processed in paraffin.
Results
A total of 194 cattle brains were examined during nine years, 136 brains from adult cattle and 58 from calves. In 115 animals no morphological lesions were diagnosed. In 77 adult samples and 2 calves a wide array of pathological lesions was diagnosed. The most important diagnosed diseases were listeriosis, rabies, malignant catarrhal fever, and several cattle of meningoencephalitis of unknown aetiology, cerebrocortical necrosis and two cases of tumours.
Conclusions
In order to be able to make an accurate diagnosis and act on time in outbreaks of important animal diseases, it is essential to know the epidemiological situation. Neuropathological examination is a substantial part of a systematic control of infectious diseases affecting the central nervous system, and should be a routine in all cases of clinical neurological symptoms.
References
- Bleem AM et al. JAVMA 1994; 204: 644
- Guarda F et al. Schweitz Arch Tierheilk 1995; 137: 101.
13
NEUROPATHOLOGICAL SURVEY OF OVINE BRAINS IN SLOVENIA
M. Pogačnik, P. Juntes
University of Ljubljana, Veterinary Faculty, Ljubljana, Slovenia
Aims
Sheep infected with certain contagious diseases of the central nervous system (CNS) may present a threat for human as well as animal health. Similar to intensified control for CNS disorders in cattle, neurological diseases in sheep are also gaining more interest. Lately, the emphasis is on transmissible spongiform encephalopathies and lentiviruses (1,2). Attention for sheep neuropathology is growing in Slovenia due to intensified sheep production during the last years and to health problems related with it. The aim of this study was to get data about brain pathology of sheep necropsied in Slovenia.
Material and methods
Brains from adult sheep (ewes and rams) and lambs were included in the study. After removing from the cranial cavity, they were checked for gross pathology lesions. If bacteriological aetiology was indicated, half of the brain was fixed in buffered formalin and half sent to the microbiology lab for further investigation. After fixation they were paraffin embedded, tissue sections stained with hematoxylin and eosin and examined with light microscope.
Results
In last ten years 597 sheep brains were examined (27,7% of all necropsied sheep), 217 from lambs and 380 from adult sheep. In 64 lambs and 109 adult sheep brains no morphological lesions were discovered. Different types and degrees of inflammatory and some other lesions were found in 23 lambs (10,6% of all lesions) and 78 adult sheep (28,7%). The main pathological lesions discovered in 130 lambs (59,9%) and 193 adult sheep (50,8%) brains were hyperaemia and/or haemorrhages.
Conclusions
Knowing neuropathology status in sheep is an important epidemiological information, and can present a basis for creation of control measures and preventive programs.
References
- Kimberlin RH. O.I.E.1996: 301
- Brodie SJ et al. Am J Pathol 1995; 146: 250
14
NATIONAL PROGRAM OF ANIMAL SPONGIFORME ENCEPHALOPATHIES MONITORING IN CROATIA
B. Šoštarić, Z. Lipej
Craotian Veterinary Institute, Zagreb, Croatia
Diseases belonging to animal spongiforme encephalopaty group accquire high attention during last few years most notably because of appearance of apparently new nosological entity known as bovine spongiforme encephalopathy. From United Kingdom were disease started, reaching epidemic proportions malady spread to several European countries. In our Country disease was neither suspected nor diagnosed. However, since Croatia is large importer of breeding cattle stock from several European countries, vigorous surveillance measures in order to prevent possible introduction of diseases belonging to spongiforme encephalopathies group has been implemented. The aim of this paper is to present the organization and monitoring program for diseases of these group.
15
SYSTEMIC MALIGNANT LYPHOMA IN AN AFRICAN TURTLE (ASTEROCHELYS RADIATA)
U. Hetzel, L. Mecklenburg, R. Kell, P. Wohlsein, W. Drommer
Institut für Pathologie der Tierärztlichen Hochschule Hannover, Hannover, Germany
Reports about lymphoid neoplasm’s in reptiles area rare. This case report describes the findings of a systemic malignant lymphoma in a twelve-year-old male Asterochelys radiate. The animal had clinically suffered from bilateral exophthalmus, fast growing neoplastic masses in the thymus and the mandibular joints region. The patient had shown inappetence and apathy for several weeks.
Aim of the study
The aim of the study was to define the neoplastic growth distribution and to characterize and identify the neoplastic cell population in Asterochelys radiate.
Methods
A full necropsy was performed immediately after euthanasia. Light microscopical, immune histochemical and electron microscopical investigations were performed on formalin, respectively glutardialdehyde fixed material. Commercial available antibodies were applied for immune histochemistry.
Results
The neoplastic cells resembled morphological mature lymphocytes and were encountered generally as perivascular infiltrates. The neoplastic cells occurred in the central nervous system, retrobulbar tissue, the oral cavity, oesophagus, trachea, lungs, thymus, spleen, liver and intestine. The cells showed round to ellipsoidal shape, eosinophilic cytoplasm and spherical nuclei with one to two nucleoli. Mitotic figures were frequent. About 40% of the neoplastic cells showed a positive cell surface reaction for CD3 antigen.
Conclusion
This study demonstrates a systemic malignant lymphoma in an adult specimen of Asterochelys radiate with a large amount of CD3-positive lymphatic cells, indicating a T-cell origin of the tumour cell population.
16
PATHOLOGICAL STUDY OF THE SPLENIC LYMPHOID FOLLICLES IN EXPERIMENTAL CLASSICAL SWINE FEVER
E. Ruíz-Villamor, F.J. Salguero, M.J. Bautista, P. Sánchez, L. Carrasco, J.C. Gómez-Villamandos
Dpt. Veterinary Pathology. Univ. Córdoba, Spain
Classical swine fever is a viral haemorrhagic disease of the swine characterised by a severe lymphopenia, with the spleen being a primary target organ for Hog cholera virus (HCV). Several lesions of the spleen have been reported and different pathogenic mechanisms for the lymphopenia have been proposed. In this paper we present the histopathological, immunohistochemical and ultrastructural changes observed in the splenic lymphoid follicles of pigs inoculated i.m. route with a virulent strain of HCV (Quillota strain, isolated in Chile). The animals were euthanized at 2,4,7,10 and 14 post-inoculation days (pid). Spleen samples were fixed in formaldehyde (4%) for histopathological and immunohistochemical techniques. Ultrastructural study was developed on samples fixed in glutaraldehyde (2.5%). Morphometric analysis of the follicular area was carried out on sections stained with H&E.
Morphometric study revealed a decrease in the splenic follicular area from 4 pid, but it was recovered from the 10th day, with a significant increase at 14 pid. Histopathological and ultrastructural studies showed that this change was associated to the apoptosis of lymphocytes observed at 2-4 pid, but not detected at the next days. At 7 pid hipocellular follicles and intrafollicular plasmatic cells were observed. Enlarged and hypercellular follicles were shown at 10-14 pid. Immunohistochemical study revealed an increased number of intrafollicular macrophages (MAC387, lysozime, ?1antitripsine), and HCV infection (gp55 HCV) of these cells and macrophages of marginal zone from 2 pid. The infection of lymphocytes was scarce and only detected from 4 pid. The dendrite cell networks did not show significant changes when were immunostained with anti-?S100.
17
CLASSICAL SWINE FEVER: IMMUNOHISTOCHEMICAL AND ULTRASTRUCTURAL STUDY OF THE RENAL GLOMERULUS
F.J. Salguero, E. Ruíz-Villamor, J.C. Gómez-Villamandos, M.J Bautista, S. Romanini, M.A. Sierra
Dpt. Veterinary Pathology. Univ. Córdoba, Spain
Renal haemorrhages, intravascular coagulation, deposits of immunecomplex and proliferation of mesangial cells have been reported as characteristic renal lesions in the different courses of Classical swine fever (CSF). In this report we present the immunohistochemical and ultrastructural changes observed in the renal glomerulus of pigs inoculated with a virulent strain of Hog cholera virus (Quillota strain, isolated in Chile) in order to determine the lesion sequence, the nature of the immunocomplex and the glomerular cellular populations infected with the HCV.
Pigs were euthanized at 2,4,7,10 and 14 post-inoculation days (pid). Kidney samples were fixed in formaldehyde (4%) for histopathological and immunohistochemical techniques. Ultrastructural study was developed on samples fixed in glutaraldehyde (2.5%).
The lesion picture observed in the glomerulus of these pigs was a moderate mesangioproliferative glomerulonephritis, in which initial changes were detected at 7 pid with proliferation of mesangium and mesangial cells. Swollen podocytes and fusion of foot processes were observed from this date. Positive immunoreaction to IgM, IgG and C1q were detected in the glomerular capillary wall and less in mesangium from 7 pid. However, subendothelial and submembranous electrondense deposits were detected only at 10-14 pid by electron microscopy. Viral antigen (gp-55 HCV) and viral structures were observed in circulating monocytes, neutrophils, glomerular endothelial cells and podocytes from 7 pid. Free viral particles and vesicles with viral particles were observed in the lumen of Bowman’s capsule.
18
IMMUNOHISTOCHEMICAL DETECTION OF PARVOVIRUSES IN TISSUES OF DOGS WITH PARVOVIROSIS – USE IN DIAGNOSTICS AND FORENSICS
T. Švara, P. Juntes, M. Pogačnik, M. Gombač, P. Hostnik
University of Ljubljana, Veterinary Faculty, Ljubljana, Slovenia
Aim
Death of an animal due to parvovirosis is a frequent cause of lawsuits, and forensic specialist often has to determine the length of the pathological process. Few references based on experimental infections were found that could help evaluating duration of infection and illness (1,2). The purpose of our investigation was:
a) to find out applicability of polyclonal antibodies for immunohistochemical demonstration of parvoviral antigen,
b) to confirm the use of immunohistochemical methods as an additional tool for determination different phases of the disease and the length since the beginning of infection.
Methods
We used routinely processed paraffin tissue sections of gut and several other tissues from twenty dogs with a known clinical history. Sections were stained with hematoxylin eosin (HE). For immunohistochemistry we used specific polyclonal antibodies against parvoviruses, raised in rabbit, and LSAB® kit (DAKO) as a visualisation system.
Results
Histological lesions pathognomonic for parvovirosis were present in gut sections stained with HE in all examined samples. Immunohistochemicaly we demonstrated parvoviral antigen in epithelial cells of intestinal villi and/or crypts and in some cases in desquamated enterocytes. Reaction product was oftenly found also in lymphocytes, macrophages, and endothelial cells of the intestinal wall capillaries and in several other tissues. In majority of our cases, we evaluated that the duration of the process was from six to nine days. This evaluation was based on gut histology only. When we used immunohistochemistry we were able to demonstrate viral antigen distributed in different compartments of intestinal wall that was characteristic for earlier phases of the disease (2), thus making time evaluation more accurate. For immunohistochemical determination of the earliest phase e.g. from day 1-3 post infection, it is necessary to include other organs in examination – thymus, lymph nodes and spleen are essential.
Conclusions
Immunohistochemical method in combination with HE staining enables precise diagnosis as well as a better estimation of infection duration.
References
- Carman PS et al. Res Vet Sci 1985, 38: 141
- Macartney L et al. Vet Rec 1984, 115: 453
19
THE DEMONSTRATION OF PERICRYPTAL FIBROBLASTS IN COLONIC INFLAMMATORY-REGENERATIVE AND DYSPLASTIC EPITHELIAL LESIONS
S. Radović, I. Selak, I. Bratović
Institute of Pathology, Sarajevo, University School of Medicine and Gastroenterology Department, Sarajevo, University Hospital Centre, BiH
Aim
The aim of the paper is definition of perycriptal fibroblasts (PCFs) distribution in inflammatory-regenerative and dysplastic changes of bowel mucosa. The relationship between the presentation of PCFs and the intensity of inflammatory-regenerative and dysplastic process in the flat colon mucosa will also be examined.
Methods
Biopsy specimens from 270 patients were examined: 74 classified as inflammatory-regenerative and 196 as dysplastic (108 mild, 58 moderate and 30 severe dysplasia). The demonstration of PCFs in biopsy specimens was investigated by immunohistochemistry using monoclonal antibody for alpha smooth muscle actin, muscle-specific actin (HHF-35), vimentin and desmin. According to the degree of PCFs development it was also evaluated separately in normal mucosa and adenocarcinomas.
Results
The PCFs were reduced in inflammatory-regenerative, and dysplatic mucosa. The reduction was significantly correlated with the intensity of epithelial dysplasia. The carcinomas tended to lack PCF network. During inflammatory-regenerative and dysplastic process in flat bowel mucosa, PCFs are generally thought to express alpha smooth muscle action, muscle-specific action, vimentin and desmin. These results established the smooth muscle differentiation features of PCFs during the transition from regenerative to dysplastic epithelial lesions.
Conclusion
These findings suggest that reduction of PCFs in dysplastic epithelial lesions may relate to the development of dysplasia in flat bowel mucosa. Reduction PCFs in dysplastic mucosa may be a marker for risk of carcinoma development in bowel mucosa.
20
NEUROLOGICAL COMPLICATIONS IN RHEUMATIC DISEASES
H.G. Fassbender
Zentrum für Rheuma-Pathologie, Mainz, Germany
The cross-linking of the muscular and tendinous system with the central nervous system as well as the autonomous nervous system is responsible for the symptomatology of the so-called rheumatic diseases: the flowing pain. Moreover, the manifestation of rheumatic diseases in parts of the central nervous system can lead to dramatic complications:
- through compression of peripheral nerves
- through involvement of the central nervous system in inflammatory processes, especially in arteritides
- through active influence of the autonomous nervous system on rheumatic tissue processes
Finally, instinctive nervous signals control the maintenance of joint structures by regulation of the joints loading. The implication of the nervous system in rheumatic diseases, by passive involvement or active participation, attracts special attention in therapeutical aspects as well.
21
IDENTIFICATION OF POSTMORTAL REMAINS FROM A MASS GRAVE IN LOVAS
G. Gusić, D. Strinović., D. Zečević, J. Škavić, Čadež, M. Kubat, V. Petrovečki
Department of Forensic Medicine and Criminology, Zagreb, Croatia
Authors described methods of identification of unknown civil war victims excavated from a mass grave in Lovas eastern Slavonija, Croatia. Autopsy was performed at the Institute of Forensic Medicine and Criminology in Zagreb
In this report authors described parameters of identifications, which are used in classic methods of identification. Close relatives confirmed each identification. In the majority of cases wives, sons, fathers and mothers did the confirmation. From 68 exhumed persons 67 were positively identified 59 of them were men and 9 were women. Identifications were so successful because the bodies, clothing, personal things, personal documents and jewellery were in a relatively good condition.
22
EPIDEMIOLOGY OF SUICIDE IN OSIJEK COUNTY
M. Ugljarević, M. Marcikić, S.Badžek
University hospital Osijek, Department of forensic medicine and pathology, Osijek, Croatia
We chose to study suicide in Osijek County focusing on recent trends and levels based on demographic characteristics.
Compilation of most descriptive statistics was performed using a standard statistical analysis computer package.
From 1986 to 1995, 329 suicide cases were reported among Osijek County residents.
Average annual suicide rate was 20.5 in100 000 county residents. Male suicides were contributed to the increased rate of suicide after 1990. There was a fairly uniform spread of suicide over the months of the year, with slight increase in the spring and the fall months, and low figure for January.
The suicide rate for males was the highest for groups age 25 to 29 and groups age 60 to 64. Among women suicide rate was the highest for groups from 60 to 64 year old.
Married and retired committed the largest number of suicides. Firearms were preferred by males and poisons by women.
There was a significant increase of suicides committed by firearms after 1990.The main reason for that increase was more guns among the population. Depression associated with physical incapacitation and intolerable loneliness was a common reason for suicide among elderly men.
23
“FOCAL” VS. “DIFFUSE” BRAIN INJURY: DOES IT REALLY MATTER?
B. Dmitrović*, S. Kurbel**, D. Jadro-Šantel***, V. Hlavka***
*Department of Pathology, Osijek University Hospital, Osijek, Croatia
**Department of Physiology, Osijek Medical School, Osijek, Croatia
***Department of Neuropathology, “Rebro” University Hospital Center, Zagreb, Croatia
There is a lot of confusion among clinicians with the strict division between deep traumatic cerebral lesions on the “focal” and the “diffuse” ones. The classical hypotheses describing the patterns of the disputable lesions are the “inner cerebral trauma” concept (Grčević N, Jakob H, 1965) and the “diffuse axonal injury” concept (Adams JH, 1980). The aims of this study were: 1. to determine the distribution of the axonal lesions on the large hemispherical brain slices, and 2. to test statistically the above mentioned hypotheses.
There were 18 cases of closed head acceleration/deceleration type injury that were examined in this research. The survival span ranged from couple of hours after sustaining the injury up to 348 days. The mean age was 36.4, ranging from 7 to 74 years of age. The hemispherical histological slides were stained with Gross-Bielschowsky silver impregnation method for axons’ visualization. There were 41 histological slides examined. The representative brain stem sections were evaluated using antibodies targeting the neurofilament subunits. Slides were divided in 1 cm wide square fields (in total 3673 fields in 41 slides) used for counting of axonal injuries. For the testing of the “inner cerebral trauma” concept, all fields were divided according to their anatomic location as “predilected” (924 fields) or “not predilected” (2749 fields). For the testing of the “diffuse axonal injury concept”, all fields were divided as “transitional” (2402 fields containing grey/white border) or “pure” (1271 fields containing only grey or only white matter). The ROC (“Receiver Operating Characteristic”) curve analysis with variable cut-off point was applied. Fields were counted as: true positive (a), false positive (b) false negative (c) or true negative (d). These numbers were used to calculate sensitivity, specificity, accuracy and (2 value. Definitive cut-off point was determined by the highest (2 value.
The axonal injury was evident in all the brain stem specimens. The axonal retraction bulbs would mostly occur in the corpus callosum and the internal and external capsule. The results of examined sample statistics confirm both observed hypotheses: “diffuse axonal injury” and “inner cerebral trauma” or “focal brain injury”. For the “focal brain injury” concept, accuracy in differentiating “predilected” from the “notpredilected” fields was 0.75 at the cut-off point of (5 lesions per “predilected” field ((2= 378.57, p<0.001). For the “diffuse brain injury” theory, accuracy in differentiating “transitional” from the “pure” fields was 0.54 at the cut-off point of (1 lesions per “transitional” field ((2= 39.15, p<0.01).
The results of the examined sample statistics confirm the both observed hypotheses: “inner cerebral trauma” and “diffuse axonal injury”. The damages are more severe in the predilected focal regions (“focal injury”) than at the cortex/white matter margins (“diffuse injury”). In the examined samples of brain tissue in cases of head injury, both types of lesions are present but with different intensities.
24
DENTAL IDENTIFICATION IN TWO BIG TRAFFIC ACCIDENTS IN CROATIA
J. Galić, Z. Kaić, V. Njemirovskij, H. Brkić, D. Zečević*
School of Dental Medicine, University of Zagreb, Zagreb, Croatia
*School of Medicine, University of Zagreb, Zagreb, Croatia
The aim of the research was to analyse the value of dental identification in the railway accident on the 30th August 1974 in Zagreb, as well as in the crash of a British and a Slovenian plane near Vrbovec on the 10th September 1976. In both cases, an odontologist has participated in the identification process.
In the railway accident, 108 victims were identified and in 5,56% of the cases teeth characteristics along with other characteristics – clothes, personal description, personal documents, fingerprints and jewellery – proved to be decisive.
Out of the victims from the plane crash, all 63 passengers and crewmembers from the British plane were identified, of which in 38,1% of the cases teeth characteristics (along with other characteristics) were decisive. From the Slovenian plane in which the passengers were mainly of German nationality, 103 victims were identified of which 29,12% due to the teeth characteristics (along with other characteristics). It is assumed that 10 bodies remained unidentified.
In the railway accident – on the contrary – the number of identified bodies on the basis of teeth was modest. The reason for that were modest antemortem data provided by friends and relatives as well as the rural origin of the victims which was characterised by the modest number of dental restorations. Significant number of dental identifications in the plane crash can be explained by the precise antemortem data provided by the odontologists. The bodies of the victims from the Slovenian plane were carbonised which resulted with the fact that in the 13,59% of the cases the identification was achieved exclusively by teeth.
The importance of dental identification at mass disasters has been proved because the teeth remain preserved in any physical conditions and in terms of evidence they have the value of fingerprints. It is also important to highlight the importance of filling out the teeth status in the dental charts because dental identification is only possible if precise antemortem data is available.
References:
- Clark DH. Int Dent J 1994;44:241.
- Brkić H et al. Acta Stomatol Croat 1998;32:409.
- Galić J et al. Acta Stomatol Croat 1995;29:197.
- Dješka S. Magistarski rad 1978.
POSTER PRESENTATIONS
p1
INTRAMEDULLARY LIPOMA OF CERVICOTHORACIC SPINAL CORD WITH INTRACRANIAL EXTENSION; CASE REPORT
A. Kogler, V. Beroš, J. Talan-Hranilović*, B. Božić
Department of Neurosurgery, “Sestre milosrdnice” University Hospital, Zagreb, Croatia
*”Ljudevit Jurak” Clinical Department of Pathology, “Sestre milosrdnice” University Hospital, Zagreb, Croatia
Intradural lipomas are rare, histologically benign neoplasms that account for less than 1% of all spinal cord tumours. Extensive intramedullary lipomas of the cervical spinal cord extending into the posterior fossa are extremely rare, with only 8 cases reported in the literature.
An intradural, intramedullary lipoma originating within the cervico-thoracic cord with significant extension into the posterior fossa is reported. The lipoma was found incidentally by CT scan at the infant’s age of 2 days. Follow-up studies by CT and MRI were performed on several occasions. Because of hydrocephalus, ventriculoperitoneal shunting was performed. After shunting and shunt revisions, surgery of lipoma was performed at the age of 2 years. At this age, the patient’s motor development, especially the lower extremity musculature, was retarded, corresponding to the age of 7-8 months. Due to the high operative risk, the patient’s parents refused to give their consent for the suggested lipoma operation earlier.
The surgery was performed to confirm the histologic diagnosis of the mass and to decompress the neural structures in the posterior cranial fossa and spinal canal. Posterior fossa craniectomy and laminectomy from C1 to T4 were performed. The lipoma was reduced by the microsurgical technique and with the use of a CO2 laser. Adequate decompression was achieved, leaving a relaxed pulsating cord and posterior fossa. Pathologic examination revealed a lipoma.
The patient’s postoperative course was uneventful. Postoperative treatment included rehabilitation that produced a good result. During the 12-month follow-up period, improvement of the motor deficit was recorded: the patient used both hands, was able to sit independently, and could walk with support.
p2
BIHEMISPHERICAL CEREBRAL GANGLIOGLIOMA AND HEMANGIOMATOUS MALFORMATION IN AN 18-MONTH-OLD-CHILD; CASE REPORT
A. Kogler, I. Radić, J. Talan-Hranilović*, B. Božić
Department of Neurosurgery, “Sestre milosrdnice” University Hospital, Zagreb, Croatia
*”Ljudevit Jurak” Clinical Department of Pathology, “Sestre milosrdnice” University Hospital, Zagreb, Croatia
In this paper a rare case of concomitant bihemispherical cerebral ganglioglioma and hemangiomatous malformation in an 18-month-old boy is presented.
Gangliogliomas are relatively uncommon tumours in childhood. They consist of a mixture of glial cells and differentiated nerve cells, and represent true neoplasms with the potential for progressive growth.
The child underwent diagnostic neuroimaging examinations (CT and MRI) for seizures and right-sided facial palsy that showed the bilaterally presence of parieto-occipital, oval, well demarcated, subcortically processes.
A biparieto-occipital craniotomy was carried out and two well-demarcated abnormalities were detected and removed in toto. Three years after the operation, the boy experienced normal psychomotor development and remained free from seizures with no therapy required. Repeat MRI showed no tumour recurrence.
Histopathologic and immunohistochemical analysis of the two processes found in our patient showed them to be a ganglioglioma and a hemangiomatous malformation, confirming the dysontogenetic origin of these alterations and association between malformations and tumour growth. The combination of ganglioglioma and hemangiomatous malformation, especially bilateral presentation of such two identical processes in a young child is very rare and was not found in the recent literature.
p3
HEMANGIOBLASTOMA OF THE CAUDA EQUINA IN A 13-YEAR-OLD BOY; A CASE REPORT
A. Kogler, B. Božić, J. Talan-Hranilović*, T. Sajko
Department of Neurosurgery, “Sestre milosrdnice” University Hospital, Zagreb, Croatia
“Ljudevit Jurak” Clinical Department of Pathology, Sestre milosrdnice University Hospital, Zagreb, Croatia
A rare case of hemangioblastoma of cauda equina in a 13-year-old boy is presented.
Spinal hemangioblastomas are rare tumours, accounting for only 1.6 to 2.1% of all primary spinal neoplasms. In one third of the cases, spinal hemangioblastomas are part of the Hippel-Lindau-disease, which is characterised by hemangioblastomas of the central nervous system, especially of the cerebellum, by vascular tumours of the retina and by visceral lesions of the kidneys and the pancreas. Hemangioblastomas at the level of the cauda equina are uncommon; only about 40 cases are known.
A boy presented with a six-month history of permanent low back pain and sciatic pain. Neurological examination showed weakness of the right leg, with hypalgesia in the right, proximal dermatome L5. Electromyography (EMG) showed a lesion of L5 and S1 roots, especially on the right side. Because a computed topography (CT) of the lumbosacral spine was negative, a magnetic resonance imaging (MRI) of the spine was performed. The MRI of the lumbar spine confirmed an intradural process at the level L2/L3.
The patient underwent surgery using microsurgical techniques. Laminectomy L2 to L3 was performed and after opening the dura, the roots of the cauda equina were identified. The yellow-white tumour was evident at the right root L5. Because no continuity of the root with the tumour was observed, a tumour of 20×10 mm was removed in toto with a resection of the nerve root. Histopathologic and immunohistochemical analysis of the tumour showed it to be a hemangioblastoma.
A few days after surgery the boy had urinary disturbances, but without earlier pain. Three months later, the boy had no neurologic deficit and a repeated MRI showed no tumour recurrence.
p4
ESTROGEN RECEPTORS IN HUMAN DENTAL PULP
S. Jukić, J. Talan Hranilović, G. Prpić Mehičić, I. Anić
School of Dentistry, Zagreb, Croatia
Estrogen stimulates endothelial cells to produce prostacyclin (PGI (2)) and nitric oxide (NO) (1,2). These effects are evidently mediated through Estrogen receptors. NO production is influenced by cyclical hormonal changes in women (3). The objective was to localise the presence of estrogen receptors in human dental pulp and to determine the difference in their expression between male and female pulp and between healthy and inflamed pulp. The pulps were collected from teeth planed for endodontic treatment due to prosthetic reason or pulpitis. The other groups were teeth extracted due to orthodontic reason. The donators ranged from age 14,5 to 46. The age, extirpation or extraction date, and for women, the first day of the last menstrual cycle were recorded. The estrogen receptors were visualised on parallel frozen sections by the mouse monoclonal antibody. Also, parallel sections were stained by hematoxilyn and eosin. Thirty of fifty-seven pulps were examined with a light microscope. In healthy pulp estrogen receptors were expressed in the odontoblastic layer and in the endothelial cells. Estrogen receptors were more reactive in inflamed pulps, either male or female. The expression of the estrogen receptors was not dependent on the age of examinees.
1. Mikkola et al., European Journal of Obstetrics, Gynaecology & Reproductive Biology 1998; 79: 75.
2. Klug et al, Pferdeheilklumde 1997; 13 : 490
3. Kharitonov et al, British Heart Journal.1994; 72: 243
p5
CHOROID PLEXUS CARCINOMA: A CASE REPORT
V. Čerina, L. Negovetić, J. Hranilović*, M. Nakić**
Department of Neurosurgery, “Sestre milosrdnice” University Hospital, Zagreb, Croatia
*Department of Pathology, University Hospital “Sestre Milosrdnice”, Zagreb, Croatia
**Department of Surgery and Haematology, Children Hospital, Zagreb, Croatia
During the 1987-1995 period, 267 children with intracranial tumours were operated on at Department of Neurosurgery.
The diagnosis of choroid plexus papilloma was made in two children, and of choroid plexus carcinoma in one child aged 45 days. Primary carcinoma of choroid plexus is a rare tumour of the central nervous system, with a higher frequency in children (up to 0,5%).
Two-dimensional transfontanellar echography and computed tomography (CT) were used for making the diagnosis. The 45-day-old male infant was operated on for obstructive hydrocephalus. Upon the placement of ventriculoperitoneal drainage according to Pudenz, removal of the tumour located in the right lateral ventricle was performed by craniotomy.
Pathology and histology of the material taken on several occasions verified the diagnosis of choroid plexus carcinoma.
During the postoperative course oncologists from the Children Hospital carried out chemotherapy, with eight therapy cycles in total.
Follow-up and CT examinations showed a relapse of choroid plexus carcinoma at the age of 18 months. The relapse of choroid plexus carcinoma spreading almost throughout the system of lateral brain ventricles bilaterally, was removed by recraniotomy. Presentation of the case of this rare choroid plexus carcinoma at the age of only 45 days confirms the reports of other authors on tumour localisation in lateral cerebral ventricles and clinical signs of increased intracranial pressure and seizures. Other authors report a average survival rate of 9 months.
p6
SUDDEN DEATH BY CRYPTOCOCCAL MENINGOENCEPHALITIS (CME)
J. Begić, J. Talan-Hranilović, I. Markoja, M. Ivanuša, J. Majetić
Main hospital Bjelovar, Bjelovar, Croatia
University hospital “Sestre milosrdnice”, Zagreb, Croatia
Aim of this article is to present a case report of a cryptococcal meningoencephalitis (CME) with a short survival period. This form of meningoencephalitis (ME) is extremely rare in our hospital. A 53-year old patient was hospitalised in intensive care of the Internal clinic in emergency service due to febrility of unknown etyiology, combined by the development of an acute psychoorganic syndrome. During the 7-day stay in the hospital under permanent laboratory and clinical treatment, aetiology of febrility and acute psychoorganic syndrome remained unknown. Patient died on the 7th day in comma et pulmonary oedema with hyperthermia of 42 degree C. Autopsy showed a macroscopically enlarged brain, weight 1450 g with signs of oedema. Histologycaly we found ME along with cryptococcus neoformans (CN) in leptomeningas and in subarachnoidal spaces, in cerebral cortex and in deeper structure in basal ganglia and capsula interna where we fo-und collections with great accumulation of CN with pseudohyphae by multiplied microglia. In parenchyma of the brain we found disseminated focus of perivascular infiltrator proof CN on paraffin blocks we used histochemical method HE, PAS, Fontana-Masson (Silver stain and modification Fantana-Masson with Mucicarmine, Fontana-Masson with Alcian blue and Fontana-Masson with Nuclear fast red) and immunohistochemical methods CD45 for T lymphocyte and GFAP for astroglia. CME is a rare form of ME, most common in patients with imunodeficiency and this is the first case in our hospital. We found no imunodeficiency in our patient, neither clinically, nor pathoanatomically. We found no changes with CN in other organs.
Literature:
- Sunhee C. Lee et al., Human Pathology, 1996; Vol 27:839
- Kyung J. et al., Journal of Clinical Microbiology, 1981; Vol 13:383.
p7
SACRAL PERINEURIAL (TARLOV) CYSTS: A REPORT OF TWO CASES
D Kovač, T Sajko, J Talan-Hranilovic*, J Hat**, Z Jajić***, M Ilej****
Department of Neurosurgery, “Sestre milosrdnice” University Hospital, Zagreb, Croatia
*Department of Pathology, “Sestre milosrdnice” University Hospital, Zagreb, Croatia
**Department of Radiology, “Sestre milosrdnice” University Hospital, Zagreb, Croatia
***Department of Reumatology and Physical medicine, “Sestre milosrdnice” University Hospital, Zagreb, Croatia
****Department of Anaesthesiology and Intensive care, University Hospital “Sestre Milosrdnice” Zagreb, Croatia
Sacral perineurial cysts are often confused with meningeal cysts and diverticula. While meningeal cysts are usually asymptomatic, perineurial cysts may be responsible for clinical symptoms.
Perineurial and meningeal cysts can be differentiated by their location, microscopically and radiologically.
Meningeal cysts develop as outpouchings from the subarachnoid space and they may be extradural or intradural and containing no neural tissue. Perineurial cysts occur at the site of posterior root ganglion with their wall, partially or even totally, composed of neural tissue. They are often multiple. On initial myelography sacral perineurial cysts do not fill with contrast medium in comparison to meningeal cysts which are visualised on.
We have described two cases of, commonly rare, sacral perineurial cysts that have been operated on at the Department of Neurosurgery.
In the first case a patient underwent surgery in another state on account of herniated disc on the L4/L5 and L5/S1 level. After performed surgery symptoms like pain and paraesthesia have recurred. MR showed the existence of a sacral perineurial cyst, and after complete cyst excision the patient made a good recovery. A surgery was performed on the second patient due to what seemed like an intradural tumour of the sacral region on computed tomography (CT) scans. Intraoperatively it was, both surgically and histologically verified as a perineurial cyst and upon its excision the patient made an excellent recovery without any neurological symptoms.
p8
SECONDARY BRAIN TUMORS AFTER THERAPY OF ACUTE LYMPHOBLASTIC LEUKEMIE
T.F. Hajnić, M. Mataija, J. Talan-Hranilović*, Lj. Cvitanović; B. Mučić
Department of Pediatrics Hematology and Oncology, “Sestre milosrdnice” University Hospital, Zagreb, Croatia
*Department of Pathology, “Ljudevit Jurak”, University Hospital “Sestre milosrdnice”, Zagreb, Croatia
Acute leukaemia is the most frequent malignant disease in children . Primarily cells can infiltrate any organ, and one of the favourable sites is brain. In order to prevent blastic infiltration of the brain, primarily in ALL, almost all of the previous protocols for curing ALL included preventive irradiation of brain in addition to combined chemotherapy. Today it is well known that aggressive polychemotherapy and irradiation of hemoblastosis and solid tumours, can cause the development of other forms of malignant diseases after 5 to 30 years.
Aim
Three patients who were radiated with 2,5 Gy during the treatment of ALL according to the previous protocols, developed secondary malignoms of the brain otherwise very rare in childhood – neuroblastoma, meningeoma and non Hodkgin lymphoma, during the period of 5 to 10 years following irradiation.
Patients
I ALL was diagnosed at the age of 3, and treated by modified Pinkel protocol. In addition to MTX taken intrathecal irradiation of the brain was performed with 2,4 Gy. Ten years after irradiation, neurologic deficits developed and CT revealed a tumour of the frontoparietal area. PHD: neuroblastoma. The patient died after two recidives (therapy: operation, polychemotherapy, and irradiation).
II ALL was diagnosed at the age of 8 months and treated according to the YU 84 protocol in combination with irradiation with 2,5 Gy. Ten years after the therapy was completed the neurologic deficits developed, clinical observation discovered a greater tumour process in the left temporoparietal area. Ablation of the tumour process, PHD: meningeoma. Control analysis: no signs of recidive or residua of tumour process.
III The third patient underwent irradiation of the brain with 2,5 Gy in combination with oncologic treatment at the age of 3. Neurologic deficits developed 7 years after complete remission. Lumbal punction was performed and cytological analysis of liquor showed non-Hodkgin lymphoma, and CT proved a solitary process of the left hemisphere meningeal. Therapy was continued abroad.
Conclusion
In these three cases we have diagnosed otherwise very rare tumours of the brain in children. The diagnoses followed the treatment of ALL according to the complex protocol, which also included irradiation of the brain. Because of this potential complications the modern protocol of treatment of ALL excludes application of high irradiation doses to the brain. Considering the age and dose of irradiation, irradiation is used only in children older than one year of age and in doses of maximal 1,2 Gy and only in the group of children with high risk and those with primarily infiltration of the brain.
| Patient | Tumour following irradiation | Type of tumour | Location | Therapy |
| I | 10 years | Neuroblastoma | Frontoparietal area | Operation, chemotherapy, radiation |
| II | 10 years | Meningeoma | Left temporoparietal area | Operation |
| III | 7 years | Non Hodkgin lymphoma | Left hemisphere meningeal | Chemotherapy |
p9
PROGRESSIVE MULTIFOCAL LEUCOENCEPHALOPATHY – A CASE REPORT
L. Pažanin, D. Tomac*, F. Knežević*, D. Pavić
KBC Rebro, Department of neuropathology, Zagreb, Croatia
*Institut za tumore, Zagreb, Croatia
Progressive multifocal leucoencephalopathy (PML) is a rarely occurring, progressive, usually fatal, demyelinating disease of the central nervous system that is associated with infection of oligodendrocytes by the neurotropic papovaviruses. The disease occurs predominantly in individuals with an immunocompromised state and impaired cellular mediated immunity due to another underlying illness. The incidence of PML is rising as a consequence of the AIDS epidemic and extensive use of irradiation and immunosuppressive therapy.
We report a case of PML in a 44 -year-old man mimicking a brain tumour. The diagnosis of PML was based on the presence of typical histologic findings in the white matter: multiple necrotic areas with complete demyelination, enlarged bizarre astrocytes, abnormal oligodendroglia with pale and ballooned nuclei containing granular inclusions. Diagnosis was confirmed by immunohistochemistry with anti-SV40 cr that revealed a lot of glial cells with strongly labelled nuclei.
p10
SPORADIC CREUTZFELDT-JAKOB DISEASE IN CROATIA – A CASE REPORT
L. Pažanin*, S. Telarović , B. Vejnović, D. Marković, L. Maračić
*KBC Rebro, Zavod za neuropatologiju, Zagreb, Croatia
Neurološki odjel opće bolnice Pula, Pula, Croatia
Creutzfeldt-Jakob disease (CJD) is a subacute spongiform encephalopathy, generally included among the group of human prion diseases. CJD occurs world-wide with an annual incidence of 1-2 cases per a population of a million. The disease is rapidly progressive and ultimately fatal. No specific treatment is available and medical management remains limited to supportive care. The process leading to neuropathological changes is still unknown.
We report clinical, neuropathological and immunohistochemical findings in a 75-year-old female patient with rapidly progressing dementia. Electroencephalography showed periodic triphasic sharp waves characteristic of CJD. Histopathological examination revealed spongiform degeneration of cerebral cortex with kuru-like plaques. Immunohistochemical staining for PrP was performed in Referent European Centre in Wien and showed synaptic pattern in cerebral cortex and basal ganglia.
p11
IS APOPTOSIS THE MECHANISM OF REGULATION OF THE PILOCYTIC ASTROCYTOMA SLOW GROWTH?
M. Ožegović, J. Talan-Hranilović, M. Belicza,
Clinical Hospital “Dubrava” and “Sestre milosrdnice” University Hospital, Zagreb, Croatia
Aim
To examine apoptosis in 15 cases of pilocytic astrocytoma: 12 benign and 3 malignant (what is extremely rare in such tumours).
There is almost no information on apoptosis in pilocytic astrocytoma in the literature. Pilocytic astrocytoma is known as a slow growth benign tumour, gradus I. Therefore we wanted to confirm if apoptosis was the mechanism of growth regulation for this tumour.
Methods
The material was treated with classical paraffin procedure, hemalaun-eosin stain, cut with microtom on cuts 5 ?m thick. To determinate the volume and numerical density of apoptotic bodies the material was treated and examined stereologically with Weibel’s test system M42, at PF of 400x.
Results
Apoptotic bodies were found neither in pilocytic astrocytoma group nor in anaplastic pilocytic astrocytoma.
Conclusion
The slow growth of pilocytic astrocytoma is not regulated by mechanism of apoptosis. This is in accordance with the data shown in the literature which point out that the mutation of p53 gene, which controls the mechanism of apoptosis is not found in pilocytic astrocytoma.
Literature:
- ELLISON DW et al., Neuropathol Appl Neurobiol 1995;21:352.
- KOKUNAI T et al., Noshuyo Byori 1995;12:15.
- PATSOURIS E et al., Clin Neuropathol 1996;15:337.
- PATT S et al., Hum Pathol 1996;27:586.
- VON DEIMLING A et al., Acta Neuropathol 1993;86:81.
p12
MENINGEAL HEMANGIOPERICYTOMA – A RADIOLOGICAL AND CLINICOPATHOLOGICAL REPORT OF TEN CASES
D. Bedek, J. Talan-Hranilović, M. Kalousek, J. Hat, M.Lovrenčić, A.Kogler , T.Hajnžić
“Sestre milosrdnice” University Hospital, Zagreb, Croatia
Aim of the study
Meningeal hemangiopericytoma is an uncommon aggressive tumour with a propensity for both local recurrence and extraneural metastases. Roentgenographic observations, clinicopathological findings and therapy of our 10 patients were analysed.
Methods used
Between 1988 and 1998 ten patients with pathohistologically confirmed diagnosis of meningeal hemangiopericytoma were under treatment in our institution. Five tumours occurred in men and five in women with median age of 29 yr. (range 4 – 69 yr.). CT scans were performed in all cases, angiography in 4, MRI in 3 and scintigraphy on 1 patient. All patients underwent surgery; seven received postoperative radiotherapy and only one chemotherapy. Because of the recurrence 4 patients were reoperated. 2 patients had intracranial, 1 intra – and extracranial metastases. To obtain pathohistological diagnosis hemalaun-eozin, Mallory, Gomori and immunohistochemical methods were used.
Results obtained
The average time of symptom manifestation (signs of high intracranial pressure and focal neurological signs) was 6 months and they were related to the tumour location. CT and MRI scanning gave no preoperative distinction between meningeal hemangiopericytoma and meningeoma. Angiograms showed a more specific picture but not sufficiently typical. Tumours were 2,5-6 cm in greatest dimension with moderate perifocal oedema. None had hyperostosis or calcification. Six tumours were supratentorial (2 with bone erosion), 2 in the posterior fossa and 2 in the lumbar spine (1 with erosion arcus and processus spinosus). All 10 unenhanced CT scans showed hyperdense tumours – 7 were heterogenous, 3 homogenous; contrast enhanced CT scans showed enhancement.
Pathohistological features of tumours agreed with common morphological patterns of hemangiopericytoma. The number of mitosis varies and is a helpful prognostic criterion. Immunohistochemically tumours were vimentin and partly S-100 protein positive. Three patients died.
Conclusions
Radiological findings (CT, MRI, angiography) of the meningeal hemangiopericytoma are characteristic but not sufficiently typical to permit an unequivocal diagnosis. Unlike in meningeomas, hyperostosis and intratumoral calcification are not present while bone erosion and necrosis are often. The diagnosis must be histologically and immunohistochemically confirmed. Radical surgical management with radiotherapy was crucial in treatment. Chemotherapy was efficient in one our patients during eight years.
p13
DISTRIBUTION OF HISTOLOGICAL CARCINOMA TYPES AND AGES OF PATIENTS WITH DIAGNOSED BREAST CANCER BEFORE AND AFTER THE WAR AGRESSION ON CROATIA IN 1991.
I. Marton, T. Leniček, M. Glasnović, M. Vučić, D. Šimunović, V. Gladić, M. Elez, M. Belicza
“Ljudevit Jurak” Clinical Department of Pathology; “Sestre Milosrdnice” University Hospital, Zagreb, Croatia
The breast cancer has lately been determined as the number one cause of cancer caused death in females of the Western regions. We recognised it as a leading mortality-morbidity issue due to data from the “Sestre Milosrdnice” University Hospital registry and National Registry for Cancer.
The goal of our study was to compare histological findings and age-dependent characteristics in 578 patients with breast cancer diagnosed at the Ljudevit Jurak Clinical Department of Pathology, Zagreb; recorded in two periods before (1980/81) and after (1997/98) the war aggression on Croatia in 1991. There were 575 women and 3 men among those patients with female to male ratio (F/M) of 191,7.
Histological type distribution in 1980 was: invasive intraductal carcinoma found in 153 (98,7%) patients and invasive lobular carcinoma in 2 (1,3%) patients. In 1981 there were 268 (99,3%) intraductal carcinoma and 2 (0,7%) lobular carcinoma. We found 69 (93,2%) intraductal carcinomas and 5 (6,8%) lobular in 1997 and 61 (92,4%) intraductal carcinomas and 5 (7,6%) lobular carcinomas in 1998. Analysis of numbers, related to early eighties and late nineties, shows a statistically significant increase of invasive lobular carcinoma from 0,9% to 7,1%.
The average age at the time of diagnosis in 1980 was 52,9 while in 1981 it was 54,0. In the 1997 average age of the patients was 60,1 and in 1998 it was 61,5, showing a noticeable increase in average age of patients between the compared two-year periods.
Conclusions given by studying changes of distribution of carcinoma types and changes of average age of diagnosis could be explained by social difficulties and stress induced by war activities in the region.
LITERATURE:
- Rosai J et al., Ackerman’s Surgical Pathology 1996: 1565.
- Parkin DM et al., Int J Cancer 1999; 80(6): 827.
- Dmitrović B et al. u Belicza M, Krušlin B (urednici), I. Hrvatski kongres patologa i sudskih medicinara , knjiga sažetaka 1996;27.
p14
MASSIVE HEPATIC NECROSIS IN YOUNG DIARRHOIC CALVES
T. Paller, M. Černe, L. Šenk, P. Juntes, A. Perdan, M. Pogačnik
University of Ljubljana, Veterinary Faculty, Ljubljana, Slovenia
Aim
This paper is a part of a research project in which pathomorphological and bacteriological studies were performed on young diarrhoeic calves. In addition to the pathological changes found on the gastrointestinal tract and on lungs, we have detected massive hepatic necrosis in the liver using pathohistological examination. There are no reports of massive liver necrosis in the literature describing neonatal diarrhoeic diseases of calves. The aim of our study was, therefore, to systematically determine the patohistological changes in the liver and to detect possible factors that might influence the occurrence of massive hepatic necrosis.
Methods
We necropsed 122 young diarrhoeic calves. We used routinely processed paraffin tissue sections of liver from 108 diarrhoeic calves. Sections were stained with hematoxilin eosin (HE).
Results
The gross lesions on the liver were not characteristic. In the majority of cases, the liver was swollen, enlarged but in some calves they were smaller and soft. Histologically, hepatic plates were completely dissociated, the majority of hepatocytes were seen as cell debris. Endothelial cells of liver sinusoids and Kupffer cells were swollen. The infiltration of mononuclear cells was seen in the interlobular interstitium. In a few cases we have detected hyperplasia of the bile canaliculi. The duration of the process in the liver was mainly evaluated as acute, in the cases of hyperplasia of bile canalicules the process was evaluated as subacute.
Conclusions
We found massive hepatic necrosis in approximately half of diarrhoeic calves. We have established that massive liver necrosis occurs more often in the diarrhoeic calves previously treated with antibiotics.
References
- Günther H et al. Arch exper Vet med 1978; 32: 273-96.
- Strombeck DR. Small animal gastroenterology. 2nd ed. London. Wolfe Publishing Ltd, 1991:574-92.
- Jubb KVF et al. Pathology of domestic animals. 4th ed. San Diego: Academic Press, 1991: 319-406.
p15
THE PRESENCE OF CRYPTOSPORIDIA SP. AT DIARRHOIC CALVES
M. Černe, T. Paller, M. Gombač, T. Švara, M. Pogačnik
University of Ljubljana, Veterinary Faculty, Ljubljana, Slovenia
Aim
Cryptosporidiosis is a disease caused by the coccidian protozoan of the genus Cryptosporidium infection usually of gastrointestinal epithelial cells. The purpose of the present report was to tabulate the locations of cryptosporidia and to choose the most suitable staining method for tissue sections and fecal smears to detect oocysts by light microscopy.
Methods
50 calves aged up to 45 days were necropsised. The clinical syndrome was chronic diarrhoea and debilitation. The tissue samples and fecal smears were collected from duodenum, jejunum, ileum, spiral colon and rectum. HE, toluidine blue, Wolbach-Giemsa and modified Ziehl-Neelsen methods were used as a staining procedure on paraffin sections and on fecal smears.
Results
Cryptosporidia has been detected in all examined parts of intestinum. All staining procedures gave positive results, which differed in number of Criptosporidia on the microscopic field. Histologic lesions of villus atrophy and marked alteration of the glandular structure of the jejunum and ileum were accompanied by cryptosporidial forms in the striated border of the villus epithelial cells and in fecal smear from rectum in 16 calves.
Conclusions
The duodenum appears to be the least favourable small intestine location, whereas middle and lower portions of the small intestine supported high concentration of the organisms. According to the staining procedure the best results for detection of Cryptosporidia on paraffin tissue sections were obtained by using Wolbach-Giemsa. On fecal smears modified Ziehl-Neelsen was effective in demonstrating these protozoa.
References
- Anderson BC, Am J Vet Res, 1984;45:1474.
- Anderson BC, Vet Pathol, 1987;24:235.
- Angus KW et al., Vet Pathol, 1982;19:67.
p16
THE OCCURRENCE OF HELICOBACTER-LIKE ORGANISMS AND OTHER BACTERIA IN GASTRIC BIOPSIES OF DOGS
M. Gombač, P. Juntes, T. Švara, M. Černe, M. Pogačnik
University of Ljubljana, Veterinary Faculty, Ljubljana, Slovenia
Aim
Association between gastritis and gastric ulcerations and gastric bacteria occurrence exists in dogs as well as in humans. The bacteria can be diagnosed histologically, bacteriologically and with other tests. The aim of our study was to examine advantages of different histological methods for revealing the presence of Helicobacter-like organisms and other bacteria in gastric biopsies.
Methods
Gastric biopsies (38 samples), measuring 0,5 to 1,0 mm, were fixed in 10% buffered formalin, embedded in paraffin, sectioned at 4 mm and stained with hematoxylin and eosin, Toluidin blue, Elliot modification of Warthin Starry silver staining and Gram stain for tissues (Goodpasture method).
Results
Results are presented in Table 1. Toluidin blue was found the best for Helicobacter-like-organisms (HLO) detection and Gram stain for other bacteria. We also, confirmed HLO with a immunohistochemical method using specific antibodies against H. pylori.
Table 1: The presence of Helicobacter-like organisms (HLO) and other bacteria found in
gastric biopsies of dogs as revealed with different staining methods.
| Bacteria present | HE | Toluidin blue | Goodpasture | Warthin-Starry |
| HLO | 16/38 | 20/38 | 17/38 | 7/38 |
| Other bacteria | 5/38 | 8/38 | 12/38 | 3/38 |
| HLO and others | 0/38 | 1/38 | 1/38 | 0/38 |
Conclusion
Toluidin blue staining was established as the best staining method for a routine detection of Helicobacter-like organisms in gastric biopsy samples of a dog. The method is fast, nonexpensive and provides very clear morphology of Helicobacter-like organisms.
References
- Jenkins CC et al. Small Animal Gastroenterology. 1997, 19: 267
- MacLachlan et al. Vet. Pathol. 1988, 25: 36
p17
CROSS-REACTIONS IN ELISA FOR DIAGNOSIS OF PARATUBERCULOSIS IN SHEEP DUE TO INFECTION WITH PSEUDOTUBERCULOSIS
M. Pislak, P. Juntes, M. Pogačnik, M. Ocepek
University of Ljubljana, Veterinary Faculty, Ljubljana, Slovenia
Aim
The objective of this study was to find out if blood sera of sheep with pseudotuberculosis contain antibodies that non-specifically bind to antigenic components of Mycobacterium avium subsp. paratuberculosis used in ELISA for paratuberculosis diagnosis.
Methods
24 sheep from a flock infected with paratuberculosis and pseudotuberculosis were necropsied. Their blood sera was taken before death during a routine examination and tested in ELISA without preabsorption of sera and in ELISA with preabsorption of sera with Mycobacterium phlei to detect antibodies to Mycobacterium avium subsp. paratuberculosis.
Results
Lesions characteristics for pseudotuberculosis were found in 24 sheep at necropsy and according to location classified into four groups. Paratuberculosis was pathomorphologicaly diagnosed in five sheep, of which three sheep were positive and two inconclusive in ELISA without preabsorption. Only one of these was positive in ELISA with preabsorption and four were negative. With analysis of ELISA results for these 24 animals we have found out that in ELISA without preabsorption 6 sera (25%) were positive, 9 sera (37,5%) were inconclusive and 9 sera (37,5%) were negative. In ELISA with preabsorption of sera with M. phlei one serum (4,2%) was positive, two sera (8,3%) were inconclusive and 21 sera (87,5%) were negative.
Conclusions
With comparison of ELISA results and results of pathomorphological examination we have determined that blood sera of sheep with pseudotuberculosis contains antibodies that bind to M.subsp. paratuberculosis antigens in ELISA. Cross-reactions occur because of common antigenic determinants of related bacteria from genera Mycobacterium in Corynebacterium (1,2,3). More cross-reactions were found in animals with pseudotuberculosis lesions located in lung and mediastnal lymph nodes. Preabsorption of sera with M. phlei antigens decreased the number of cross-reactions considerably and with that non-specific results.
References
- Dubash K et al. J Vet Diagn Invest 1995; 7: 347-51.
- Milner AR et al. Res Vet Sci 1987; 42: 140-4.
- Pepin M et al. Vet Rec 1987; 120: 236.
p18
PREDICTIVE POWER OF KI 67 ANTIGEN EXPRESSION IN UVEAL MELANOMAS
A. Krvavica, J. Talan-Hranilović*, M. Belicza*
General Hospital, Zadar, Croatia
*University Hospital “Sestre milosrdnice”, Zagreb, Croatia
Uveal melanomas are made of two distinctive cell types, spindle and epitheloid, with differing clinical implications. These melanomas are known to be less aggre-ssive malignant tumours comparing to the cutaneous type of same cell characteristic (1).
The intent of this short study was to investigate expression of Ki-67 immunocomplex in uveal melanoma cells, examine if intensity of expression correlates with levels of invasion, which would result in prognostic significance.
19 uveal melanomas were tested for the presence of Ki-67 protein by indirect peroxidase antiperoxidase method and monodonal antibody for Ki-67 protein. The intensity of the reaction is marked by a semi-quantative method.
All samples of epitheloid melanomas showed strongly positive Ki-67 reaction. Spindle B cells, which are a transi-tional type between epitheloid and spindle A cells, show greater variation in Ki-67 expression, from negative to moderately positive.
Literature:
- Brown DC and Gatter KC. Monoclonal antibody Ki-67 its use in histopathology. Hystopathol 1990; 17:489
- Sternberger LA et al. J.Hystochem cytochem 1970; 18:315
- McCormick D, Chang H, Hobbs C et al. Histopathol 1993; 22:355
- Resnick M et al. Hum Pathol 1996; 27:234
- Costa MJ et al. Hum Pathol 1996; 27:274
- Weidner N, Moore DH, Vartanian R. Hum Pathol 1996; 2:2337
p19
ISOLATED BRAIN LANGERHANS´ HISTIOCYTOSIS – A CASE REPORT
K. Žarković, G. Jurić, V. Nesek, J. Papa, Z. Heinrich, V. Hlavka, J. Paladino
Department of Neuropathology, Clinical Hospital Zagreb, Zagreb, Croatia
Introduction
Involvement of the brain by Langerhans´ histiocytosis (LH) is seen in one quarter of patients with systemic disease. Isolated brain involvement is rare and usually includes hypothalamic and pituitary lesions. Often CNS lesions are multifocal and may arise in a synchronous or metachronous fashion. A primary lesion of CNS LH is present occasionally in the non-hypothalamic region.
Clinical details
A 34-year-old female had symptomatic epilepsy. CT scan showed a tumour in the left temporal region, measuring 4 cm in diameter, which looked like meningeoma. Post-neurosurgical pathological diagnosis of the biopsy showed chronic inflammation. No diagnostic or therapeutic treatment was performed after surgery. Four years later patient suffered from right side limb weakness, motoric dysphasia and 2 weeks of progressive loss of appetite and depression. CT scan and MRI showed two lesions measuring 2 cm in diameter in the white matter of the right frontal region and in the lateral left diencephalon region and parietal region. A stereotactic biopsy was performed.
Neuropathology findings
The initial biopsy yielded eight needle cores of tissue which showed cerebral tissue infiltrated by a lot of macrophages (S-100, CD 68 and ?1-antichymotripsin immuno-positive), T and B-lymphocytes, plasma cells and microglial cell clusters. Lot of reactive astrocytes and blood vessels proliferated. Perivascular inflammatory infiltration was composed predominantly of T-lymphocytes. In one out of eight cores of tissue Langerhans´ giant cells with folded nucleus were noticed. Electron microscopy of de-paraffinated tissue was performed, but penta-laminar Birbeck’ s granules were not found.
Conclusion
It is important, but also, very difficult to make a diagnosis of Langerhans´ hystiocytosis in a small biopsy sample, because of numerous histologic variations that could look like unspecific inflammation (incl. viral infections), Non–Hodgkin lymphomas, other granulomatos inflammatory processes with epitheloid proliferation, or like other hystiocytosis variations.
p20
P53 AND BCL-2 EXPRESSION IN EXTRAHEPATIC BILE DUCT CARCINOMAS
A. Kopjar, M. Vučić, B. Šeni, H. Hochstaadter, H. Čupić, M. Belicza, B. Krušlin
Varaždin, Zagreb – Croatia
Aim
The aim of the present study was to analyse the expression p53 and bcl-2 in the extrahepatic bilc duct carcinomas (EHBDC).
Materijal and methods
We analysed tumour 13 patients with EHBDC’s diagnosed at “Sestre milosrdnice” University Hospital in the 1990-1996 period. There were 10 female and 3 male patients ranging in age from 56-94 years. In 8 patients only palliative treatments was performed. Five patients underwent surgical resection. Histologic analysis revealed adenocarcinoma in all cases. Formalin fixed, paraffin embedded tumour tissue of these five patients was cut at 5mm, deparaffinized, and stained with an antibody to p53 and bcl-2 (Dako, Denmark). Alkaline phosphatase/antialkaline phosphatase method was used. The intensity of immunostaining was graded semiquantitively and designated with (-) for no staining, (+) for 10% positive cells, (++) for 25% positive cells and (+++) for more than 25% positive cells.
Results
Histopathologic examination showed p53 expression in all cases: in 2 tumours the reaction was weakly positive (+), and in 3 tumours strongly positive (+++). We found weakly positive immunostaining for bcl-2 in 2 tumours, moderately positive in one tumour and strongly positive in 2 tumours (Table).
Conclusion
We conclude that EHBDC’s are uncommon tumours and the results of tretment at present are rather poor. It seems that p53 and bcl-2 are present EHBDC indicating their possible role in the development of these tumours. Assessmrnt of clinical and pathological impact of these genes on EHBDC requires evaluation in larger studies.
Case |
bcl-2 |
p53 |
1 |
+ |
+++ |
2 |
+++ |
+ |
3 |
+ |
+ |
4 |
++ |
+++ |
5 |
+++ |
+++ |
p21
DEVELOPMENT OF BLOOD VESSELS IN HUMAN FOETAL TESTIS
D. Ježek, Lj. Banek, A. Hittmair*, M. Kos, H. Rogatsch* and G. Mikuz*
Institute of Histology and Embryology, Institute of Pathology Medical School University of Zagreb
* Institute of Pathology, University of Innsbruck
Aim of the study
In the testis, blood vessels are situated in the interstitial compartment. They contribute to the hypothalamus-hypophysis-gonadal interaction and are vital for the normal function of seminiferous tubules. There is a lack of data on blood vessels in the foetal human testis. Therefore, the aim of our study was to investigate the development of blood vessels at different maturation stages (gestational weeks) of the human foetal testis.
Material & methods
Foetal testes (15-36 weeks of gestation) were obtained during the routine pathological section of miscarried children. Immediately after section, the testes were fixed in Bouin’s fluid for 24 hours. Following fixation, tissue specimens were subjected to the usual histological procedure and embedded in paraffin. 4 µm sections were made by a rotation microtome. Several sections were stained with haemalaun and eosin and some were chosen for immunohistochemistry (IHC) in order to visualise blood vessels. For this purpose, antibodies to factor VIII (Von Willebrand factor, F VIII) and alpha-smooth actin were used. IHC results were quantified using a multipurpose Weibel’s test system with 42 points at a magnification of x400.
Results
A fine network of blood vessels was noticed in the testicular interstitial compartment at each investigated stage of development. Large blood vessels could be observed only in septula and the mediastinum testis area. All types of blood vessels (i.e. arteries, veins and capillaries) expressed F VIII. However, the expression of alpha-smooth actin was restricted only to arteries and veins that possess smooth muscle cells in their wall. Morphometric analysis demonstrated a gradual increase in number and volume of all types of blood vessels over the investigated period of gestation.
Conclusion
Our study showed that, during development, blood vessels of the human foetal testis increase in number and volume. It could be presumed that such a developmental dynamics of vessels is necessary for the secretion of foetal testosterone and the hypothalamus-hypophysis-gonadal interaction/s.
p22
MORPHOLOGICAL PROPERTIES OF TESTICULAR BIOPSY IN A VASECTOMY PATIENT
D. Ježek, S. Škrablin*, I. Krhen**, I. Kuvačić* and Z. Mareković**
Institute of Histology & Embryology, Gynaecology and Obstetrics Clinic** Urology Clinic, Medical School University of Zagreb
Aim of the study
The testicular biopsy is performed in the case of severe oligoasthenospermia and azoospermia. Azoospermia is caused by many known and unknown factors. One of the factors could be a congenital absence of vas deferens or vasectomy. Herein, we present one case of a vasectomy patient.
Materials & methods
The patient consulted an andrologist for the assisted reproduction procedure (in early fifties). At 28 y. of age he was subjected to vasectomy. Before the vasectomy, he fathered one child. The bioptic material was divided into several small parts with an average dimension of 1 mm3. Six parts of the biopsy were frozen for the further assisted reproduction procedure, whereas two parts (originating form each testis) were immediately fixed in 5% glutaraldehyde (in 0.1 M phosphate buffer). Postfixation was performed with 1% OsO4 and tissue specimens embedded in Durcopan. Semi-thin sections were made by a Reichert ultramicrotome and stained with 1% toluidine blue.
Results
Biopsy was composed of 15-20 seminiferous tubules and testicular interstitial tissue. Most seminiferous tubules had a normal diameter (120-180 *m) and a regular architecture of the lamina propria. The seminiferous epithelium was composed of spermatogonia (A and B), spermatocytes, early spermatids, late spermatids and sperms. In some seminiferous tubules the spermatogenic cells were disorganised and desquamated in the tubular lumen. Sertoli cells of such tubules had an increased number of vacuoles and lipid droplets. Despite disorganisation and desquamation of spermatogenic cells, late spermatids and sperms were also occasionally found in the above mentioned tubules. In one or two tubules macrophages of the interstitial tissue have entered the tubular lumen and were phagocyting several sperms (spermatophagia).
Conclusion
Morphological analysis of the testicular biopsy in the vasectomy patient demonstrated spermatophagia in some tubules as a response of immunocompetent cells to the vasectomy. However, based on the histological examination, the frozen bioptic material could be used for the assisted reproduction procedure.
p23
GIZZEROSINE INDUCED HISTOPATHOLOGICAL LESIONS IN BROILER CHICKS
M. Tišljarl, Z. Grabarević2, B. Artuković2, Z. Šimec2 , P, Džaja3, V. Herak-Perković4
l Poultry Centre, Croatian Veterinary Institute, Zagreb, Croatia
2 Department of General Pathology and Pathological Morphology
3 Department of Forensic Veterinary Medicine, Veterinary Faculty, University of Zagreb, Zagreb, Croatia
4 “PLIVA” Research Institute, Zagreb, Croatia
Gizzerosine is a well-known toxic substance spontaneously produced during the heating process of fish meal. Because of its action on H2-receptors of histamine it stimulates gastric juice secretion and induces gizzard erosions (GE) (Okazaki et al., 1983; Masumura et al., 1985). Its pathological effects on other organs have not yet been described.
In this work the pathological effects of gizzerosine on different chicken organs were investigated. The ROSS broiler chicks were divided into three groups: group A received 100% of non-medicated commercial mash for broiler chicks. For group B 50% of commercial mash was replaced with unheated fish meal (0.5 ppm gizzerosine) and in group C with heated fish meal (1.15 ppm gizzerosine) during an experimental 7-day period. Fourteen chicks from each group were sacrificed every day during one week. Necropsy did not reveal notable differences between treated groups. Samples of gastrointestinal, cardiovascular and lymphoid organs, of brain and kidney were collected for histopathological analysis; fixed in 10% formalin solution, embedded in paraffin, sections of 5 ?m thickness were cut on a microtome and stained with hematoxilyn-eosin stain.
Histopathological findings predominantly developed in gastrointestinal organs (proventricular mucosal degeneration; intestinal mucosal and submucosal edema with/without multiple vacuoles; proventricular and intestinal submucosal fibroplasia; gizzard cuticle erosions).
The most prominent were intestinal lesions; their severity increased towards the end of the trial, especially in group C.
References
- Okazaki et al ., 1983; Agric. Biol. Chem. 47: 2949.
- Masumura et al., 1985; Poult. Sci. 64: 356
p24
MARTEILIOSIS IN MUSSEL (MYTILUS GALLOPROVINCIALIS) AS POTENTIAL HAZARD IN CROATIAN SHELLFISH PRODUCTION
S. Zrnčić, D.Oraić, B. Šoštarić
Croatian Veterinary Institute, Zagreb, Croatia
As a Mediterranean country Croatia has a millennium-old tradition of shellfish farming. Domestic oyster (Ostrea edulis) and mussel (Mytilus galloprovincialis) are traditionally cultivated in several places along the Adriatic coast, especially Malostonski bay and Limski bay.
Global shellfish industry has been affected by various very serious epizootics questioning profitability of further production. One of the most important is marteiliosis caused by protistan parasites of the phylum Paramyxea, genus Marteilia. However, Marteilia species were observed in a variety of hosts including oyster (O.edulis), mussels (M.edulis and M. galloprovincialis), cockles (Cardium edule) and clams (Tapes spp.). The parasite develops mainly in the epithelium cells of the digestive gland and is associated with emaciation of the host and exhaustion of its energy reserves (glycogen).
Diagnosis is established by examination of the smears of infected organs or by histologic means stained sections of the digestive gland.
During the monitoring of shellfish health status, marteiliosis in mussel was observed.
In this paper the description of clinical and histopathological study of the case is presented.
p25
PATHOMORPHOLOGICAL PICTURE OF TULAREMIA IN EUROPEAN BROWN HARE
Ž. Mihaljević, Z. Lipej, B. Šoštarić, M. Žurić.
Croatian Veterinary Institute, Zagreb, Croatia
Tularemia as a zoonotic disease has been well recognized for a long time. Causative agent is a small, pleomorphic gram-negative coccobacillus.
Francisella tularensis. There are several forms of the disease in humans, varying from localized skin lesions, more or less generalized lymphadenitis, pneumonia or granulomatous ocular lesions. Although many animal species are harboring causative agent, brown hare (Lepus europeus europeus, Pallas) is the most frequent source of human infection in our country. Despite of relatively high occurrence in hare, pathoanatomical lesions are seldom well observed due too frequent post mortal changes of cadavers submitted for examination. Pathomorphological picture of tularemia in hares recently observed in our Institution is presented in this work. Most notably on hare, hepatosplenomegaly with generalized lymphadenopathy is observed. Necrotising hepatitis, characterized by disseminated necrotic foci, although generally described has been observed only on rare occasions.
p26
DIAGNOSTIC PROBLEMS OF LISTERIOSIS IN SHEEP
Z. Lipej, B. Šoštarić, Ž. Mihaljević, M. Žurić
Craotian Veterinary Institute, Zagreb, Croatia
Listeriosis, an infectious disease caused by Gram-positive microorganism Listeria monocitogenes is most, in sheep frequently manifested clinically an pathomorphologically in two separated forms, namely as septicemia with consecutive abortion or somewhat specific type of encephalitis. That later form of the disease is highly suspected on the basis of spoiled silage feeding history and almost diagnostic by clinical behavior of affected animals. Adding to that evidences, pathoanatomical and characteristic histopathological findings, the disease is practically pathognomonic. However in our experience bacteriological intentions to isolate causative agent either from silage or brain tissue presenting obvious histopathological changes are seldomly successful. In these work we present c. history data coupled to pathomorphological findings of several cases diagnosed as listeriosis at our Institute during last two years. Causative agent was was not bacteriologically determined in any if these cases.
p27
A NON-INFECTIOUS ENTEROPATHY INDUCED IN CARP (CYPRINUS CARPIO L.) BY STARVATION
F. Božić, E. Srebočan, Z. Kozarić
Veterinary Faculty, Zagreb, Croatia
A four-week study was conducted to shed light on the effects of starvation-induced stress on local immune factors and histological changes in the fish gut mucosa. In this study we found that carp starved for four weeks developed enteropathy, comprising villus atrophy, stratum compactum hyperplasia and a significant decrease in intestinal intraepithelial lymphocyte (i-IEL) numbers. These changes were associated with a dense cellular infiltrate into the lamina propria. In addition, we found here an increase in goblet cells (GC) number in starved carp gut. However, there were qualitative changes in GC mucins, neutral mucins becoming predominant in starved carps suggesting functional adaptation of GC to the changed conditions in the gut throughout mucus hyperproduction and specific composition of mucins as well. These results indicate that the pathobiology of starvation-induced i-IELs decrease, matching GC proliferation and inflammatory cells homing to the gut, could be an adaptive mechanism to sress which probably enable compensatory immune surveillance in the fish gut mucosa.
p28
AELUROSTRONGYLOSIS IN CATS
Ž. Grabarević, S. Ćurić, B. Artuković, Z Šimec, K. Ramadan
Faculty of Veterinary Medicine, Zagreb, Croatia
Aelurostrongylus abstrusus (A.a.) is a parasite with a indirect life cycle of a lung parenchyma in the cat. Its predominant sites are terminal bronchioles and alveolar ducts. In this study we have examinated two population of cats. The first group consisted of feral cats (n=50) in the rural areas of County of Zagorje (Nortwestern Croatia), and the second group encounted cats from the urban area of City of Zagreb. For the second group data was compilated from fecal examination from the Department of Parasitology and the histopathologycal and gross pathological finding on cats lungs at the Department of General Pathology and Pathological Morphology Faculty of Veterinary Medicine, University of Zagreb. In the first group histopathological examination was carried out and the A.a. was diagnosed in 22% (n=11), and in the second group A.a. was diagnosed in 0,4% (n=2) of the fecal examination and in eight year period (1990-1998) and 3,9% (n=5) on histopathological examination of cats’ lungs also, in the same eight year period. In most of the cases histopathological examination revealed alveoli, bronchioli and alveolar ducti filled with eggs and larvae of A.a., disruption of alveolar septae, cellular infiltrate (mostly eosinophyls, macropahges, lymphocytes and few multinucleated giant cells. Frequently, peribronchial cell infiltration with the same cellular type, a hyperplasia and hypertrophy of smoothe muscle of pulmonic arteries. This study also brings literature overview on Aelurostrongylus abstrusus.
p29
THYROID CANCER IN HOSPITAL’S THYROID DISEASE REGISTRY
D. Šimunović, I. Cvjetko, B. Krušlin, H. Čupić, M. Lechpammer, Z. Kusić, M. Belicza
“Ljudevit Jurak” Clinical Department of Pathology “Sestre Milosrdnice” University Hospital, Zagreb, Croatia
Aims
By using our registry and analysing, both pathological and clinical data, of our patients, we searched for patterns or distractions in epidemiological and clinical behaviour of thyroid cancer during last 20 years.
Methods
Thyroid cancer registry was filled with data from post-surgery biopsy findings in the University Hospital, clinical data both preoperative and postoperative and then connected to National Registry of Cancer for follow up data.
Results
Thyroid cancer registry comes with 433 patients in total pool of 3815 thyroid disease patients contained in Hospitals Thyroid Disease Registry. Papillary cancer is most common with 264 (61%) cases and female to male ratio (F/M) of 3,3:1. Follicular cancer is next with 72 (16,6%) cases, F/M of 2:1, medullary carcinoma comes with 35 (8,1%) cases and F/M ratio of 1,3:1. Anaplastic cancer with 32 (7,4%) cases and F/M ratio 1,9:1 formed the smallest group. Survival rates are as follows: 5-year survival for papillary cancer is 97%, follicular cancer 93% and for the medullary carcinoma 78%. In order to gain advanced information considering age distribution we compared average age at diagnosis in two different 2-year periods (1980-81 versus 1997-98).
Conclusion
Sex distribution in our registry is as quoted in literature. Follicular cancer is on the lower border of distribution for countries with abnormal iodine uptake what is in correlation with the presence of endemic goiter in Croatia, but with decreasing incidence from 15 in 1980 to 3 in 1997 and none in 1998. Survival rates are as quoted. By comparing two different 2-year periods we found that average age at diagnosis of follicular cancer increased from 45,5 years (1980-81) to 61,3 years (1997-98).
References
- Kusić Z. et al., Endocrinol Invest 1996; 19: 204.
- Teppo L. et al., Eur J. Cancer 1998; 34: 2248.
- Hundahl SA et al., Cancer 1998; 83: 2638
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HOSPITAL’S THYROID DISEASE REGISTRY 80-98
I. Cvjetko, V. Gladić, M. Elez, D. Šimunović, M. Glasnović, T. Leniček, M. Vučić, B. Krušlin, H. Čupić, Z. Kusić, M. Belicza
“Ljudevit Jurak” Clinical Department of Pathology “Sestre Milosrdnice” University Hospital, Zagreb, Croatia
After completing the Hospital’s Thyroid Diseases Registry, our aim was to analyse any regular scheme of thyroid disease appearance, thus giving us powerful predictive mechanisms for future patients.
We used biopsy findings that were obtained after thyroid surgery from 1980 to 1998. Other relevant clinical data of each patient was also, added.
Total of 3815 patients’ data is present in our registry. Out of those 2403 (63 %) were patients with non-tumour disorders, while 1412 (37%) patients had thyroid tumours. Goiter is the most common, total of 2021 patients with female to male ratio (F/M) of 6,8. Morbus Basedow was found in 156 patients with F/M 8,1 and average age at diagnosis 39,8 years. 138 patients had Hashimoto thyreoiditis, average age 48,4 and F/M of high 14,3 Adenomas were most common in patients with tumours making total of 979 patients with F/M of 6,1 and average age at diagnosis 44,5 years. Thyroid cancer was present in 433 patients with papillary cancer as the most common with 264 cases, followed by follicular cancer with 72, medullar with 35 and anaplastic with 30 cases. Further more, we compared the average age for non-tumour disorders in two 2-year periods, using years 80/81 and 97 /98.
Valuable data obtained from the registry will enable better prediction of thyroid diseases and thus make prevention or cure more efficient, especially when combined with other clinical data ( levels of calcitonin, thyreoglobulin etc.)
References
- Pagliara S et al., Ann Ist Super Sanita 1998; 34(3): 417.
- Lind P et al., Thyroid 1998; 8(12): 1179.
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IMMUNOHISTOCHEMICAL DETECTION OF P53, PCNA AND KI-67 EXPRESSION IN ORAL LICHEN PLANUS IN CORRELATION WITH CLINICOPATHOLOGICAL FINDINGS OF THE ORAL MUCOSA
D. Biočina-Lukenda, A. Pirkić, A. Cekić-Arambašin
School of Dental Medicine, University of Zagreb; Clin. Department of Pathology, ”Sestre Milosrdnice” University Hospital, Zagreb, Croatia
Purpose
The aim of this pilot study was to examine p53, PCNA and Ki-67 expression in oral epithelium correlated with the severity of clinicopathological alteration of oral mucosa in oral lichen planus (OLP).
Method
Biopsies of the oral mucosa from ten patients with clinical picture of the OLP of various clinical degrees of inflammation, hyperkeratosis and erosions were immunocytochemicaly examined for p53 and for proliferative antigens (PCNA and Ki-67) expression using the ABC method after previous pre-treatment in a microwave oven.
Results
Marked inflammation as well as hyperkeratosis of mostly middle to strong degree was clinically observed in all patients. In cases with erosive OLP, size of erosions varied between 2 to 8 mm. The reaction of examined antigens was mosaically expressed in the epithelium; it was intranuclear and focally prominent in groups of cells. Proliferative antigen expression was also, observed in the epithelial intercellular bridges, but not for p53. Marked cytotopographic differences were observed among examined antigens. P53 was expressed in the basal cell layer exclusively, PCNA in the basal, parabasal and spinous cell layers, and Ki-67 only in the basal layer of the oral epithelium. The intensity of antigen expression varied accordingly to the degree of epithelial dysplasia.
Conclusion
The intensity of immunohistochemical reaction of p53, PCNA and Ki-67 expression was more prominent in the oral epithelium of the patients with severe clinicopathologic status of the OLP.
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THE EFFECT OF BPC 157 AND GENTAMYCINE ON REPAIR MORPHODYNAMIC OF ALKALI INDUCED CORNEAL ULCERS IN RABBITS
B. Pirkić, Ž. Matičić, Ž. Grabarević, A. Pirkić
Clinic for Surgery, Orthopedics and Ophtalmology of Veterinary Faculty, Department of General Pathology and Pathological Morphology of Veterinary Faculty; Clin. Department of Pathology,”Sestre Milosrdnice” University Hospital, Zagreb, Croatia.
Purpose
In this work we have shown the dynamics of histological changes in alkali wounded rabbit cornea during therapy with BPC 157, gentamycine-atropine combination and saline in different groups.
Methods
Corneas of 30 Chinchilla Bastard rabbits were wounded using 30 application of 2N NaOH through a 9 mm wide plastic tube firmly attached to the eye. Therapy lasted for 17 days and considered of BPC solution in first group, 0,3% gentamycine and 1% atropine solution in second and saline in the third group, each drug administrated locally. Groups consisted of 10 animals each, and because of monitoring changes during the treatment, extirpation of the eyes was performed on days 3, 6, 9, 14 and 17. In addition, fluoresceine staining was performed every day. Enucleated eyes were fixated in buffered 4% paraformaldehyde during 24 hours, and then cut through the equator and through the lesion margin, and embedded in paraffin.
Observations
Histologically, initial lesion resulted in necrosis of epithelial and endothelial layer, loss of Bowman capsule, oedema and fiber dissolution of substantia propria, along with vacuolar degeneration and kariorhexis of keratocytes. Descemet membrane was the only undamaged cornea structure. Monitoring the dynamics of histological changes during 17 days showed progressive development of retrocorneal membrane with iridocorneal synechia, together with the presence of vascularized scaring connective tissue on the site of corneal lesion, as two most prominent complications. Neovascularization of various degrees was markedly present in the limbal region. Both complications persisted through the whole treatment period, but development of retrocorneal membrane was significantly lower in the group that received BPC 157 treatment, comparing with other two groups, and also that group did not develop any iridocorneal synechia.
Conclusion
We have shown that the development of scaring connective tissue along with iridocorneal synechia, being the most important complications after alkali wounded corneas, were significantly reduced after being treated with BPC 157.