ACADEMY OF MEDICAL SCIENCES CROATIA
"Ljudevit Jurak" Clinical Department
of Pathology "Sestre milosrdnice" University Hospital Zagreb
and Department of General Pathology
and Pathological Morphology of the Veterinary Faculty University of
Zagreb
9th LJUDEVIT JURAK INTERNATIONAL
SYMPOSIUM ON COMPARATIVE PATHOLOGY
June 5-6. 1998. ZAGREB, HOTEL SHERATON
CROATIA
BOOK OF ABSTRACTS
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A.
Pathological
morphology of human and animal diseases
Patološka
morfologija humanih i animalnih bolesti
Prostate
cancer: Stage prediction by preoperative biopsies
Mikuz G. Rogatsch H
Institute of Pathology,
Innsbruck, Austria
The tumor stage and the grade of malignancy are the most important
parameters forthe prognostic and therapeutic assessment of the prostate
cancer at the moment.To distinguish localized (pT2) from capsule-transgressing
(pT3) carcinomas before starting therapy would, therefore, be an important
goaLThe clinical methods used for this purpose either have a low accuracy
or can only be perFormed with a very expensive equipment available only
in few specialized hospitals.Transrectal core needle biopsy is however,
perFormed in all patients with elevated PSA and/or other symptoms suspicious
for prostate cancer. Several authors tried to determinate the tumors' stage
by morphological and morphometrical analysis of such biopsies. We analyzed
90 prostatectomy specimens and matching needle core biopsies. One half
of the patients (N=45) had localized carcinoma (pT2) and the other half
of prostatectomy specimens showed an extraprostatic tumor extension (pT3).The
Gleason score perFormed on core needle biopsies showed only a minimal difference
between the two stages. A simple method for predicting the extension of
prostate carcinoma is to count the number of biopsies which contain carcinoma.
The method works, however, only if at least 6 biopsies are perFormed. The
linear length of carcinoma (in mm) in needle core biopsies can also be
used as predictive parameter. In our material pT2 carcinomas showed an
average length of 3,4 ~ 2 mm in the biopsies, whereas the average length
of carcinomas with extracapsular extension was 20 ± 5 mm (p<0,000 I
).The favorite method used by a number of authors seems to be the measurement
of the relative carcinoma volume (vol%) of carcinoma in the core needle
biopsies. In our material the mean relative volume of carcinoma tissue
in the needle core biopsy of patients with pT2 stage in prostatectomy specimen
was 4,9 ± 2,7 vol% pT3 carcinomas in prostatectomy material showed an average
relative volume of 30,1 ± 23% in the biopsy (p<0,0001 ). A method previously
used in our laboratory was the assessment of the microvessel density in
biopsy specimens pT2 carcinomas showed a significantly lower vessel density
than pT3. In our opinion the measurement of the relative volume of carcinoma
in core needle biopsies fulfills all these criteria in the best possible
way. Moreover,the method is not time consuming and does not require special
equipment - a pathologist needs no more than a simple ruler to measure
the length of the carcinoma as well as of the single biopsy chips.
Indications,
technique and complications of prostatic needle biopsies
Reljić A,
Spajić B, Kraljić I
Department of Urology
"Sestre milosrdnice" University Hospital, Zagreb, Croatia
Prostatic adenocarcinoma is the third most common malignant tumor in
Croatian male population and represents a major public health problem.
About 85% of prostatic adenocarcinomas are diagnosed in advanced stage.Two
years ago, a multidisciplinary approach to the early diagnosis of this
disease has been introduced at the Urological Clinic,"Sestre milosrdnice"
University Hospital in Zagreb.The aim of this project was to increase the
proportion of patients diagnosed in earlier stages of the disease.We present
our current protocol for establishing the indication for prostatic needle
core biopsy. The experiences on I 34 patients concerning the preoperative
treatment, bioptic techniques and complications of the procedure are shown.
We conclude that prostatic needle core biopsies are necessary for the
early diagnosis of prostatic adenocarcinoma.This procedure could be performed
in local anesthesia with minimal complications.
The
relationship between clinical and pathohistological fndings in prostatic
needle biopsies
Reljić A,
Marković-Puač Lj, Spajić B, Tarle M, Čupić H, Kraljić I, Krušlin B, Belicza
M
Departments
of Urology, Pathology and Oncology and Nuclear Medicine
"Sestre
milosrdnice" University Hospital, Zagreb, Croatia
Aim Prostatic needle core biopsy is confirmative
method for prostate cancer diagnosis. During a two-year period at Clinic
of Urology, University Hospital "Sestre milosrdnice", 250 patients were
examined for prostate carcinoma (PC).The aim of this study was to analyze
clinical and pathohistological findings in prostatic needle core byopsies
in regard of early diagnosis of the disease.
Methods The population consisted of ambulatory
patients over 50 years of age with lower urinary tract symptoms. All patients
were examined according to protocol for early diagnosis of PC. The patients
underwent digitorectal examination (DRE) and serum prostate specifc antigen
(PSA-Tandem R) measurement. lndication for needle core biopsy were positive
DRE and/or PSA value above 4 ng/mL.These. patients underwent transrectal
ultrasound examination (TRUS).All visible leasions of peripheral prostatic
zone were biopsied and sextant biopsy was taken. If no leasion was detected,
random sextant biopsy was perFormed.
Results According to above mentioned
criteria I 34 of 250 patients underwent biopsy (53,6%). DRE was positive
in 67 (50.0%) andTRUS in 70 (52.0%) patients.The PSA level was below 4
ng/mL in 20 of biopsied patients ( I 4.9%), between 4 and I 0 in 66 (49.2%)
and above I 0 ng/mL in 48 (35.8%) of patients. Prostatic cancer was found
in 39 (29. I %), high-grade PIN (HGP) in I 9 ( I 4. I %) and benign prostatic
hyperplasia (BPH) in 76 (56.7%) of patients. PC with low Gleason score
(2-4) was in 17 PC (44.0%), with Gleason score 5-7 in 8 patients (20.0%)
and with high Gleason score in 14 (36.0%) patients. In cases with well
diferentiated cancers PSA was 47.7 ng/ mL (range 5.8-105.0), moderatelly
differentiated cancers 68.8 ng/mL (range 17.9-131.0) and in poorly differentiated
PC I 8.4 ng/mL (range 0.02-49.8). Average PSA for PC patients was 43.8
ng/mL, for HGP I0.9 ng/mL and 7.6 ng/mL for BPH patients.Volume of the
prostate with cancer was roughly 25.0% lower than for BPH. Of 39 detected
PC I 2 patients (30.7%) had indication for radical prostatectomy and clinical
understaging was 58.3%.There was upgrading of 56.0% of PC in radical prostatectomy
specimens comparing with needle core biopsies.
Sensitivity of DRE was 92.0%, specificity 67.0%,
positive predictive value (PPV) and 54.0% and negative predictive value
(NPV) 96.0%.Almost identical results were obtained forTRUS. HGP was visualised,
by means ofTRUS, in 45.0%. PC was found 5 I .0% and HGP in 10.0% of hypoechoic
peripheral zone lesions. For PSA value above 4.0 ng/mL sensitivity was
95.0%, specifoity only I 9.0%, and PPV 32.0%. For PSA value above I 0.0
ng/mL, sensitivity was 69.0%, specificity 78.0%, and PPV 56.0%.
Conclusions No single test (DRE, PSA,TRUS)
had diagnostic capabilities for PC and HGP DRE and PSA should be considered
as first line tool for early diagnosis of PC. Although we have biopsied
almost every DRE positive and virtually all patients with PSA above I 0
ng/mL,there is enough space for reducing the number of unneccessary biopsies.
Tumor neoangiogenesis
in prostatic carcinoma
Volavšek M, Mašera A, Ovčak Z
Institute of Pathology, Medical
Faculty, Ljubljana, Slovenia
Introduction Tumor neoangiogenesis has proven its prognostic
value in malignant tumors of different organs. The association with disease
progression and metastatic spread has also been shown in prostatic carcinoma
after radical prostatectomy.
Aim The aim of our retrospective study was to disclose its significance
in incidental prostatic carcinoma aftertransurethral resection (TURP).We
examined all ICP diagnosed at the Institute of Pathology, Medical Faculty
in Ljubljana in years 1985-89.
Methods To determinate how microvessels correlate with the degree
of tumor differetiation determined histopathologically and with Gleason
score, stage of disease, and the time of survival in at least 6 years after
diagnosis, we counted microvessels within the initial carcinoma in 68 patients.
Microvessels were higlighted by immunostaining endothelial cells for factorVlll-related
antigen and counted in a x200 microscope field (0,8012 mm2) in the most
active areas of neovascularisation, Neoangiogenesis and degree of tumor
differentiation were then correlated between primary tumors and rebiopsies
in I 0 patients. Degree of tumor differentiation was also correlated with
the time the disease progression occured.
Results Higher microvessel counts were associated with
lower degree oftumor differentiation (p=0,005), Gleason score (p=0,001
), stage of disease in primary tumors (p=0,003), and survival of the patients
(p=0,05).The time of disease progression in I 0 rebiopsied patients showed
association with lower level of tumor differentiation (p=0,004; 0,006).
Microvessel counts were significantly higher in re-biopsies comparing to
initial tumors (p=0,006), but no differences could be observed in the degree
of differentiation.
Conclusion Our results showed that the determination of
microvessel density in TURP biopsy specimens of ICP retains its prognostic
value already demonstrated in radical prostatectomy specimens of clinical
prostatic carcinoma.
A
study of transvesical prostatectomy: Retrospective-prospective analysis
of 230 Cases
Božić Z,
Kraus O, Duančić V
Department
of Urology "Sestre milosrdnice" University Hospital, Zagreb, Croatia Institute
for Cardiovascular Prevention and Rehabilitation, Zagreb, Croatia
A total of 230 men undergoing transvesical prostatectomy
(TP) for benign prostatic hyperplasia (BPH), with glands estimated to weigh
more than 60 g, were analyzed retrospectively and followed-up one year
afterthe operation.Age, symptoms, test results, histopathological fmdings,
postoperative complications and additional operations were evaluated. Reported
dysuric symptoms were obstructive in 47,8%, irritative in 26,5%, and combined
in 20,4% of the patients. Hematuria was the main symptom in 5,2% of the
patients. BPH was histologicaly determined in 87,0%, incidental carcinoma
of the prostate in 4,3%, and various types of prostatitis in 8,7% cases.
Complications of TP had a lethal outcome in 2,2%, bleeding requiring further
surgical procedure in 6, I %, epididymitis in 4,8%,wound infection in 2,6%,
urethral stricture in 0,9%, incontinence in 0,9% and postoperative urinary
retention without urethral obstruction in 2,2% of the patients,The most
frequent reason for reoperation was bleeding in 66,7% cases. Uretheroscopic
measurement of the length of the prostatic urethra, measurement ofthe post-micturition
residual volumen and intravenous urography cannot be used as a diagnostic
criteria in the selection of patients forTP Digital rectal examination
of the prostate and cystoscopy retain their diagnostic significance, while
ultrasonography offers the best diagnostic possibilites. Significant drawbacks
ofTP are high rates of postoperative bleeding and early postoperative lethality.
Renal
carcinoma: An epidemiological assessment in two periods of time. (Trieste,
1985-86 vs 1994-95)
Bussani R,
Pavletić N, Silvestri F
Institute
of Pathological Anatomy - University of Trieste, Italy
Renal cell carcinoma (RCC) is more frequently
encountered in older patients and results in significant mortality.The
relatively poor outcome may be due to frequent late clinical presentation
and resistance of the tumour to the most current forms of adjuvant therapy.The
poor prognosis of RCC underlines the importance of determinating the real
incidence of the lesion and the necessity for finding some useful prognostic
parameters.The purposes of our study are to identify incidence patterns
of RCC over two periods of time in the province ofTrieste, and to evaluate
the biological evolution of these lesions. One hundred-fourteen new cases
of RCC were histologically diagnosed among 29,323 biopsies and 5,656 autopsies
performed between I 985-86 at the Institute of Pathological Anatomy of
the University ofTrieste. In the period 1994-95, among 36,200 biopsies
and 4,600 autopsies, a histological diagnosis of RCC has been made in 99
cases.The average annual incidence was I 3.6 in males and 8.2 in females
in the first period oftime, versus I 6.3 and 5.5 in the two sexes respectively
in the second period of time. A RCC has been diagnosed only at autopsy
in 56 out of the I I 4 cases evidenced between I 985-86 and in 45 out of
99 cases diagnosed between 1994-95. In both periods of time age specific
rates of RCC (bioptic and at autopsy diagnosis) increased proportionally
with age, reaching the peak incidence in subjects aged 75 and over. On
the contrary age specific rates of RCC clinically discovered during life
increased only until the 65-74 decade in both periods examined.0ut ofthe
58 patients with RCC diagnosed during life between I 985-86, 42% died from
the renal tumour, 4% decreased with the tumour, 27% decreased without signs
of the neoplasm and 27% are alive and well. Out of the 54 patients with
RCC diagnosed during life between I 994-95, only I 3% died from renal cancer.
As for the RCC evidenced at autopsy, about 40% of the patients died from
their tumour in both periods of time. Ninety percent of the patients pathologically
staged as T I -T2 healed, versus 43% of those stagedT3 and more.The 5-year
adjusted survival rate for patients with RCC graded as G I -G2 was 78%,
while the survival curves of subjects with RCC graded as G3-G4 was 85%
within the fifth year of follow-up.The results from our study indicate
that up to today about 50% of RCC are evidenced only at autopsy.Tumour
staging seems to be a significant prognostic factor, while nuclear grade
was found to be a non important criterion when considering the outcome
of patients with RCC.
Papillary
renal carcinoma (clear cell type) arising in the excretory urothelial district.
A very unusual feature. Morphohistopathological findings
Bussani R,
Pellegrino M, Silvestri F
Institute
of Pathological Anatomy, University of Trieste, Italy
Clinical features A 37-year-old
man was admitted to the hospital for gross hematuria and frequency The
cystoscopy was negative. An abdominal computed tomografic scan discovered
a mass arising in the calycopielic district ofthe right kidney. The clinical
impression was for a transitional carcinoma ofthe escretory tract. A nefroureterectomy
was performed.
Gross pathology The kidney weighed
178 grams, and its dimensions were I 2x6.5x3.5 cm; the resected ureter
measured 7.5 cm.The intermediate calyx and a part of the renal pelvis were
flfed by a white-greyish slightly papillary tumour. Another small roundish
white mass was evident in the subcalyceal district, without apparent continuity
with the greater lesion: No lesions in the renal cortex and medulla or
in the ureteral district were evident.
Microscopic pathology The tumour
was composed of solid sheets of clear cells interspersed with abundant
thin-walled blood vessels; sometimes abortive papillae and glomeruloid
structures were evident.The tumour cells exhibited diffuse PAS-positivity
and reacted with cytokeratin AE I , AE3, vimentin and Ulex-Europeus, whereas
there was not positive staining for CEA and S I OO.The tumour macroscopically
resembling a transitional carcinoma but characterised by microscopic and
immunohistochemical features typical of a clear cell renal carcinoma could
be considered as a sort of hamartomatous lesion.
Correlation
between vimentin expression and nuclear grade in renal cell carcinoma
Kopjar A, Dimanovski
J, Stanić G, Krušlin B, Belicza M
Department of Urology
University Hospital "Sestre milosrdnice", Zagreb, Croatia
Department of Clinical
Pathology `Zjudevit Jurak" Clinical Hospital "Sestre milosrdnice", Zagreb
Croatia
Aim To analyze the correlation between the expression of vimentin
and nuclear grade of renal cell carcinoma (RCC).
Methods In this study, tumor tissue from 20 patients was
analyzed. There were 7 male and I 3 female patients with RCC conf ned to
the kidney (pT I , pT2), ranging in age from 36-80 years (mean 60,6 years).Thirteen
tumors measured 5 cm in diameter and 7 were <5 cm.Two tumors were papillary
and I 8 showed non-papillary pattern. Six tumors were composed of granular
cells and I 4 of clear cells. Formalin fixed, paraffin-embedded tumortissue
was cut at 5 Nm, deparaffnized, and stained with a monoclonal antibody
to vimentin (Dako, Danemark).The intensity of immunostaining was graded
semiquantitatively and denoted as (-) for no staining; (+) for weak; (++)
for moderate; and (+++) for strong reaction. Nuclear grade was determined
according to Fuhrman et al. Am J Surg Pathol I 982;6:655.
Results There were 4 tumors with nuclear grade I , 3 with grade
2, I I with grade 3, and 2 with nuclear grade 4. Histopathological examination
showed vimentin expression in eight cases (40.0%), while I 2 tumors (60.0%)
were negative. Positive reaction for vimentin was observed in all four
tumors grade I , none of three tumors grade 2, four of I I tumors with
grade 3, and in none of 2 tumors with nuclear grade 4. In our study there
was no correlation between the expression of vimentin and nuclear grade
of RCC.
Conclusion Our study shows a lack of correlation between
the expression of vimentin and nuclear grade in renal cell carcinomas.
Further studies on a larger series of tumors are obviously needed.
The
influence of pH on Ki-67 antigen retrieval in the renal cell carcinoma
Mašera A, Staniša O,
Ovčak Z
Institute of Pathology
Medical Faculty, Ljubljana, Slovenia
Introduction Heat based antigen retrieval (HBAR) method using
different retrieval solutions is evaluated by immunohistochemical demonstration
of Ki-67 antigen in the renal cell carcinoma. In various tissues the optimal
conditions forthe antigen retrieval (AR) should be determined by HBAR in
solutions with different pH values (1,2).
Material and methods Tissue sections from 20 tumor specimens
were treated for I 5 min at 97 C in the laboratory microwave oven.Three
retrieval solutions were used: 0,05 M citrate buffer pH 6,0; 5mM EDTA/
NaOH pH 8,0 and Target Retrieval Solution (TRS) pH 9,9 DAKO. The automated
labeled streptavidin biotin-DAB reaction were performed on the DAKOTechMate
500.Working dilutions ofthe monoclonal antibody MIB-I (DIANOVA) were 1:150,
1:300, 1:600, respectively.
Results Optimal immunohistochemical reaction was achieved perForming
HBAR in citrate buffer with antibody dilution I:150. Using the EDTA/NaOH
and TRS the best results were obtained by antibody dilution I :600. Morphometric
analysis showed significant higher number of positive nuclei of tumor cells
after HBAR in EDTA/NaOH orTRS comparing to the HBAR in citrate buffer.
Conclusions Ki-67 antigen couldn't be demonstrated in formalin
fixed-parafn embedded specimens,therefore AR method has to be used. In
renal cell carcinoma we found the following conditions yielding the best
results:HBAR in EDTA/NaOH pH 8,0 andTRS pH 9,9, respectively.The addition
of EDTA as a chelating agent for calcium ions enhances the results of HBAR
(3).
Literature 1) Shi S-R et al., J Histochem Cytochem I 997; 45:327.
2) Taylor CR et al., Hum Pathol I 994; 3:263
3) Morgan JM et al., J Pathol I 997; I 82:233
Use
of immunohistochemistry in the diagnosis of renal angiomyolipoma
Mašera A, Zidar N, Ovčak
Z
Institute of Pathology,
Medical Faculty, Ljubljana, Slovenia
Introduction Renal angiomyolipoma (AML) is a rare benign
tumor of mixed mesenchimal origin.The diagnosis is based on the recognition
of the three components - mature adipose tissue, smooth muscle cells, (which
can occasionally be polymorphic) and characteristic blood vessels with
thickened walls, but without elastic lamellae. However, all three components
are not always present, especially in small biopsy samples. In such cases
AML can be misinterpreted for a sarcoma.
Aim To determine whether immunohistochemistry might be helpful
in distinguishing AML from other mesenchymal tumors especially tumors of
the smooth muscle origin.
Methods Twenty cases ofAML were stained immunohistochemically
with antibodies (Ab) against vimentin, smooth muscle actin, desmin, HMB-45
and NKI/C3. Control group consisted of leiomyoma, leiomyosarcoma and lipoma
(five cases in each group).
Results Afl cases of AML stained diffusely with Ab against
vimentin, smooth muscle actin, desmin and NKI/ C3, and only focally with
Ab against HMB-45. Leiomyomas and leiomyosarcomas stained with Ab against
vimentin, desmin and actin, but not with Ab against NKI/c3 and HMB-45.
Lipomas did not stain with any of the Ab used.
Discussion The results of our study indicate that immunohistochemistry
can be useful in the diagnosis of AML. Staining with anti HMB-45 and NKI/C3
distinguishes AML from benign and malignant smooth muscle cell tumors,
and it might be useful in small biopsy samples in which all three components
ofAML are not present. Finally, our study showed that NKI/C3 stains smooth
muscle cells ofAML diffusely and not only focally as HMB-45 and,therefore,
should be regarded as a better markerfor AML than HMB-45.
Congenital
malformations of genitourinary system in the I 0-years periods: An autopsy
study
Kos M, Ilić-Forko D,
Babić M, Nola M, Mayer D, Tabain I, Jukić S
Department of Gynaecological
and Perinatal Pathology, Medical School University of Zagreb, Zagreb, Croatia
Aim To determine the incidence and types of urogenital malformations
on our autopsy material in period from I 988- I 997.
Method Retrospective analysis of autopsy records and paraff
n sections when available.
Results 7672 autopsies of spontanously or medically aborted
fetuses, stillborn and liveborn infants that died in the neonatal period
were perFormed at our Department. In 17. I % ( I 32 cases), a malformation
of urinary/genital system was diagnosed. Bilateral renal agenesis was diagnosed
in I 6.6% (22/ I 32); in 68. I 8% it was isolated, in 3 I .8% combined
with other malformations, mostly anal and rectal (57, I 4%). Unilateral
renal agenesis was diagnosed in 9.8% ( I 3/ I 32). It was unexpected and
isolated in I 5.4% and combined with other malformations in 84.6% cases.
Renal hypoplasia was diagnosed in 6.8 I % (9/ I 32) of cases; it was an
isolated f nding (except for pulmonary hypoplasia) in I I.I % while it
was combined with other malformations in 88.8%.Anomalies of position and
form were noted in I 7.4% (23/ I 32); in 73,9% the shape was anomalous
(94. I % were kidneys fused at midline) while anomaly of position was diagnosed
in 26. I % (6/23). Fused kidneys were an isolated malformation in 30.43%
(7/23 cases). Renal cystic disease was found in 23.48% (3 I / I 32); dysplastic
kidneys comprised 58.06%;ADPKD 9.67%,ARPKD and renal cortical microcysts
6.45` o each and in I 6. I 3% the type of the cystic disease could not
be determined. Renal cystic disease was an isolated f nding in 29.03%,
and in 70.96% it was combined with other malformations (in 50% those of
CNS). In 9.09% of renal cystic disease combined with other anomalies (2/22),
Meckel Gruber syndrome was diagnosed. Prune belly syndrome was diagnosed
in 6.8 I % (9/ I 32); pathological changes of ureters (stenosis, duplications,
dilatations with consequent hydronephrosis) in 12.9% ( I 7/ I 32) and malformations
of the bladder and urogenital sinus 2.3% (3/ I 32) of cases. Isolated anomalies
of the genital system were diagnosed in 4.5% (6/ I 32) of the cases.
Conclusion The results of this study differ somewhat from
the results of other authors, mostly from Western Europe and American continent.
It remains to be clarified whetherthe differences are due exclusively to
ethnic causes or to different approaches in material evaluation.
Mast
cell in testicufar biopsies of infertile men
Banek Lj, Schulze
W Goluža T, Hittmair A, Pezerović-Panijan R, Ježek D
Medical Paculty Uniu
of Zagreb, Zagreb, Croatia
Mast cells of the human testis are occasionally observed within the
gonadal interstitium. In some cases of idiopathic male infertility, these
cells are known to increase in number. Since there is a lack of data on
mast cells in the cases of male infertility, the aim of our study was:
a) to characterize these cells according to their location within the testis,
b) to determine their number and volume and c) to correlate this data with
the status of spermatogenesis (evaluated by the modified )ohnsen score
/1-10/).Testicular biopsies (Epon embedded) were divided into the control
and infertile group.The blocks were serially cut in order to obtain semithin
and ultrathin sections. Semithfn sections were stained with I % toluidine
blue and observedlmeasured underthe binocular light microscope (Carl Zeiss).Within
the biopsy, mast cells could easily be recognized by their oval (or a little
bit elongated) shape and typicai numerous dark-stained granules.The cells
were found on two characteristic locations; a) in the vicinity of seminiferous
tubules or within the lamina propria itself ("peritubular-" mast cells)
and b) in the proximity of blood vessels and Leydig cells of the interstitium
("interstitial" mast cells). Stereological analysis indicated a signifcantly
increased number of mast cells in the group of infertile patients when
compared to control biopsies.A close association between the low score
and the increased number of mast cells was found.The data in our study
indicates a significant involvement of mast cells in the male infertility.
Literature: Hedger MF Rev Reprod (1997), 2:38 Yamamoto M et
al. Acta Urol Japon (1 994), 40:541 NagaiT et al. Fertility Sterility (1992),
57: 1331 Hashimoto J et al. Urologia Internat (1988) 43:129 Agarwal S et
al. Int J Fertil (1 987) 32:283 Haider SG et al. Andrologia (1 985) 17:532
Expression
of KiM I P, testosterone and Inhibin in testes of infertile men
Ježek D Lebo A, Hittmair
A, Rogatsch H, Schulze W
Medical Faculty Univ,
of Zagreb, Zagreb, Croatia
Within the testis, various cell populations interact by numerous paracrine
factors.This is especially true for Sertoli and Leydig cells. (n addition,
recent studies on testicular macrophages have shown that these cells, situated
within the testis interstitium, could affect testosterone production of
Leydig cells. Therefore, the aim of our study was to investigate the expression
of KiM I P I (macrophage marker), testosterone and inhibin in testicular
biopsies of infertile men.Testicular biopsies were divided into 2 groups:
8 control biopsies and 36 biopsies that displayed various degrees of hypospermatogenesis.Volume
of each analyzed testis was determined by ultrasound or orchidometer. Spermatogenesis
was evaluated using a modified Johnsen score ( I - I 0) (De Kretser and
Holstein, I 976). Cells positive to KiM I P I , testosterone and alfa-subunit
of inhibin were measured by the Weibel multipurpose system with 42 test
points.Testis of infertile patients displayed a significantly lower testicular
volume and score when compared to controls. Morphometric (stereological
analysis) showed a significant increase in number and volume of macrophages,
testosterone-producing cells (Leydig cails) and inhibin-producing cells
(Sertoli and some Leydig cells) per mm3 of the testicular tissue when compared
to control values.When the same variables were analyzed per entire organ
(absolute number and volume of positive cells identified by immunohistochemistry)
no signifcant difference between the two groups could be established (except
for number of inhibin-positive Leydig cells which were significantly increased
in the group of infertile patients). A significant correlation was found
between macrophages, testosterone- and inhibin-producing cells,testis volume
and score. Moreover, a positive association between the expression of inhibin,
testosterone and macrophages was also found. This data suggests that Sertoli
cells, Leydig cells and macrophages could act as a paracrine unit within
the testis.
Immunohistochemistry
of spermatocytic seminoma
Vodovnik A, Mašera A,
Ovčak Z
Institute of Pathology,
Medical Faculty Ljubljana, Slovenia
Aim To evaluate a pattern of immunohistochemical reactivity
in the spermatocytic seminoma.
Introduction Spermatocytic seminoma (SS) is a rare tumor
arising in testis only. Immunohistochemistry of SS was rarely investigated
so far, exposing discrepancies in cytokeratin and NSE positivity of tumor
cells (1).
Material and methods Three cases of SS were studied. Patients
were 51 ,61 , and 63-year-old. In all patients an orchiectomy was perFormed.
Immunohistochemical reaction by streptavidin-biotin complex method on alpha-fetoprotein,
hCG, wide spectrum keratin, cytokeratins 13, 17, 18, 19 and 20, vimentin,
LCA, EMA, CEA, PLAP NSE, S-100 protein, chromogranin A, synaptophysin,
Ki-67, and PCNA was performed.
Results Tumor cells in all three cases showed positive
immunohistochemical reactions on NSE, vimentin, PCNA, and Ki-67. PCNA and
Ki-67 immunopositivity was strong to moderate in the nuclei of tumor cells.
Cytoplasmic positivity of NSE and vimentin was moderate in all tumor cells.
No positive reactions on PLAP and cytokeratins were found. Other immunohistochemical
markers revealed negative results as well.
Conclusions Recentfy,the PLAP and NSE immunoreactivity
in classic seminoma was claimed by some authors as exclusive ( I ). According
to our findings, the usefulness of NSE as a discriminator between classic
seminoma and SS seems to be questionable. On the ground of wide array of
cytokeratins used in our study, we tend to deem these markers as helpful
in the differential diagnosis.
Literature. I . Cummings OW et al., Hum Pathoi 1994; 25:54.
Carcinoma
of the penis - A case report
Lenz D, Čačinović V
Našice District Hospital,
Našice, Hrvatska
We report a patient 52-years of age who presented with undefined complaints
in the region of the glans of the penis. Local finding suggested phymosis.
Clinical examination revealed small induration measuring up to I cm while
inguinal lymph nodes were enlarged.The patient refused surgery and was
seen 8 months later with advanced disease. Total penectomy with inguinal
lymphadenectomy was perFormed. Pathohistological analysis revealed a tumor
measuring up to 6,5 cm. Histological finding was well-differentiated squamous
cell carcinoma infiltrating urethra. The patient is at present alive and
without recurrence.
CYTOLOGY
AND QUANTITATIVE CYTOMORPHOLOGY IN NEPHROGENIC ADENOMA
Wiener H G, Remkes G
W Mian C, Pycha A, Susani M
Departments of Clinical
Pathology and Urology Uniu Vienna, Austria
Predominantly found in transplant patients the incidence of nephrogenic
adenomas increases. Although considered a metaplastic lesion of the transitional
epithelium, malignant transformation cannot be excluded.The aim of the
presented study was to describe the value of cytology and quantitative
cytomorphology in the follow up of these lesions.
Methods Seven patients with primarily transurethrally
resected, histologically proven, nephrogenic adenomas were followed for
two years. In summary, I 5 voided urines and bladder washings were available,
Cytological diagnosis was done on cytopsin preparations of voided urine
and bladder washings. Static image analysis was done on Feuglen stained
bladder washings. Based on dual parameter morphometry using the DNA parameter
2cDl and a shape descriptor a risk factor for bladder cancer was estimated.
Results Recurrent or persisting lesions were found at
five cystoscopic controls of three patients. Cytological evaluation of
voided urine reported either cellular features negative for atypia and/or
malignancy (n= I 2) orthose summarized as atypia were found (n=3). Static
image analysis resulted in estimation of a low (n= I 0) or intermediate
(n=2) risk for bladder cancer.Three specimens had to be considered insufficient
for image analysis. Recurrent/persisting lesions were removed by transurethral
resection and were proven as nephrogenic adenomas.
Conclusion The evaluated morphological data stress the
benign biological behavior of nephrogenic adenoma and prove cytology and
quantitative cytomorphology as most useful parameters in the follow up
of these lesions.
Lymphocytic adenohypophysitis
simulating pituitary adenoma
Talan-Hranilović J, Čupić H, Gnjidić
Ž, Hat J
Department of Clinical Patkology
`Ljudevit Jurak" "Sestre milosrdnice" University Hospital, Zagreb Clinical
Department of Radiology "Sestre milosrdnice" University Hospital,
Zagreb Clinical Department of Neurosurgery "Sestre milosrdnice" University
Hospital, Zagreb, Croatia
Lymphocytic adenohypophysitis (LAH) is a rare autoimmune inflammatory
process ofthe pituitary gland occurring predominantly during the peripartal
period. LAH can cause pituitary expansion and hypopituitarism presenting
with features suggestive of a pituitary adenoma.We report a case of a 39-year-old
woman who developed bitemporal hemianopsia and artralgic pain in the small
joints during the late stage of her second pregnancy. Five months after
the delivery visual field defects withdrew, but serum hormone analysis
revealed hypothyroidism and increased levels of PRL. Magnetic resonance
imaging showed an intrasellar mass with suprasellar extension measuring
I 6x I 2mm, that closely resembled pituitary adenoma. A transphenoidal
approach was used to remove the patients' sellar lesion and all of the
abnormal tissue was removed. Histopathological examination of the tumor
showed an extensive mononuclear infiltration of the adenohypophysis tissue
by plasma cells and lymphocytes forming follicles with germinal centers.
Neurohypophysis was uninvolved, morphologically normaLThe immunoperoxidase
technique showed difFuse presence of immunoreactive prolactin within surviving
adenohypophysal cells. Immunohistochemistry revealed CD3 and CD8 positiveT
cells at the periphery of lymphatic follicles and in the interfollicular
areas. CD 20 positivity was observed in the follicular centers. Diffuse
positivity for CD45 was observed throughout the lesion. Our data suggest
that the disorder is related to cell-mediated immunity as well as humoral
immunity.The question of surgical treatment, of this rare pathological
entity, still remains open.
Literature: 1. Hashimoto K et al. Endocr J ( I 997), 44; 1-1
0 2. IshiharaT et al. Endocr J (1996), 43; 433-40 3. Hartmann I et al.
Clin Neuropathol (1 996),15; 234-239 4. Presmann EK et al. J Reprod Med
( I 995), 40; 251-259
Correlation between
eosinophil infiltration and hyperactive response to destilled water chalenge
In chronic rhinosinuitis
Kalogjera L, Glumbić I, Zurak K,
Baudoin T, Rudnički M, Krušlin B
Departments of Otorhinolaryngology
and Patholog;y "Sestre milosrdnice"
University Hospital, Zagreb,
Croatia
It is well documented that non-isotonic aerosols induce hyper-reactive
response in nasal mucosa, which can be expressed as the increase in nasal
resistance, quantity of nasal secretion or concentration of inflammatory
mediators in nasal lavage fluid.The relationship between non specific response
to different stimuli and mediators to eosinophil proteins concentration
in tissue and nasal lavage was studied on different nasal provocation models,
but the relationship between eosinophil inf Itration in sinonasal mucosa
and nasal responsiveness to distilled water challenge has not been elucidated.
This study was designed to analyze the relationship between eosinophil
(eo) infiltration in sinus mucosa of the patients suffering from chronic
rhinosinuitis and their response to distilled water provocation.
Material and methods.
A group of I 5 patients ( I 0 male, 5 female) were provoked with the
inhalation of I 0 ccm of destilled waterTheir response to such a provocation
was estimated through the succeeding increase in nasal resistance, Nasal
resistance was measured by active anterior rhinomanometry with the computer
coupled rhinomanometerAtmos PC200 (Atmos, Lenzkirch, BRD). Basal resistance
was calculated as the mean ofthree measurements before provocation, and
following the provocation with simultaneous inhalation of I 0 ccm of distilled
water; 3 measurements were performed within fve minutes.All patients were
operated in the period of one month after nasal provocation. Samples of
nasal polyps, and mucosa from the ethmoid and maxillary sinuses was fixed
in the 10% neutral formalin, embedded in paraffine and stained with hemalaun
eosine. Eo and neutrophil leukocyte inf Itration was analyzed in I 0 high
power fields (400x) per sample and patients were divided in three groups
according to the level of eo infiltration in at least one sample: I .)
less than I 0 eo per HPF; 2.) I 0 to 20 eo per HPF ; and 3.) more than
20 eo per HPF Neutrophil infiltration was not recorded in any ofthe samples.
Statistical analysis ofthe data was perFormed by using the Mann Whitney
Wilcoxon test.
Results In 6 patients eo infiltration exceedect 20 eo per HPF
(group 3.); I patient had eo inf Itration of I 0 to 20 eo per HPF; and
8 patients had less than I 0 eo per HPF. One patient was excluded from
the I st group, as he had no evidence of sinus disease and was operated
for hypertrophic rhinitis only As the 2nd group was too small for statistical
analysis, the patients from the 2nd group were merged with the 3rd group.
Mean basal resistance for the 2nd and 3rd group was 0,25 Pa/cm3s- I and
for the I st group 0, I 8 Pa/cm3s- I . Mean nasal resistance following
provocation was 0,37 Pa/cm3s- ( for the 2nd and 3rd group and 0, I 9 Pa/
cm3s- I for the I st group. Mean increase in 2ndand 3rd group was 34 %
and for the I st group 26% (insignificant). Our results confirm previous
results of Klementsson ( I 99 I ), which have not demonstrated correlation
between ECP levels in nasal lavage and hyper-reactive response to metacholine
provocation considering increased secretion to provocation.The basal and
provoked resistance's in 2nd and 3rd group are significantly higher than
in the I st group.
Conclusions Considering these findings, we can conclude that
that patients with chronic sinuitis and eo inf Itration in the sinus mucosa
of more than 10 eo per HPF have higher nasal resistance than patients with
chronic sinuitis without marked eosinophil infiltration, but their responsiveness
to distilled water provocation is not significantly higher.
FEMALE ADNEXAL
TUMOR 0F PROBABLE WOLFFIAN ORIGIN
Tomić S, Alujević A, Pešutić-Pisac
V Vuković S
Clinical Hospital Split, Split
Croatia
We report a case of a 39-year old women with adnexal tumor, which arose
within the leaves of the broad ligament, connected to the left ovary and
the Fallopian tube.The patient remains alive, without evidence of the disease,
one year after left salpingoophorectomy, without subsequent adjuvant therapy.
Histologically, the tumor cells were arranged in three different patterns:
closely packed tubules, solid and sieve like. Cells had uniform round and
oval nuclei and mitoses were rare. Immunohistochemical profile of the tumor
was similar to that of Wolffian duct remnants. They co-expressed cytokeratin
and vimentin and lacked EMA reactivity, in contrast to tumors of M,Ilerian
origin which usually express EMA. Although the majority of these tumors
are benign, a few cases have shown malignant potentiaLTherefore, such tumors
should receive careful follow-up for possible recurrence and /or metastasis.
Literature: 1) Scully RE. Female adnexal tumor of probable Wolff
an origin. Cancer I 973, 3 I :67 I -677.
2) Daya D. Malignant female adnexal tumor of probableWolffan origin.Arch
Path Lab Med I 994, I I 8:3 I 0-3 I 2.
3) Rahilly MA. Female adnexal tumor of probable Wollfan origin: clinicopathological
and immunohistochemical study ofthree cases. Histopathology I 995,26:69-74.
The
malignant mixed tumor of the gallblader (Adenosquamous carcinoma with fibrosarcoma).
A case report
Alujević A, Janković
S, Družijanić N
Clinical Hospital
Split, Split, Croatia
A case of adenosquamous carcinoma with fibrosarcoma of the gallblader
is described. Adenosquamous carcinoma was characterized by the presence
of squamous epithelial and glandular components.Atypical glands were found,
among the areas of squamous cell carcinoma, which also contained squamous
epithelium with signs of cell atypia.The sarcomatous component was presented
by interwoven fascicles of spindle and fusiform, poorly differentiated
ceIIs.The clinical and X-ray examinations of the patients revealed a chronic
cholecystitis.
References: I ) Suster S. Adenosquamous carcinoma of the gallblader
with spindle cell features. Histopathology I 987, I I :209-2 I 4
2) Kuaskina G. Adenosquamous cancer of the gallblader. Arkh Patol I
980, 42:57-59
LOCAL
IMMUNE RESPONSE IN DYSPLASTIC EPITHELIAL LESIONS IN FLAT BOWEL MUCOSA
Radović S, Selak I, Bratović
I
Institute of Pathology,
Sarajevo University School of Medicine
Gastroenterology Department,
Sarajevo University Hospital Center, Sarajevo, Bosnia and Herzegovina.
Aim Dysplastic changes of flat bowel mucosa have been investigated
immunohistologicaly to determine the cellular immune response.The relationship
between the phenotype and localization of the lymphoreticular cells and
prognosis of the precancerosis has been assessed.
Methods Biopsy specimens from 170 patients were examined:
74 classifed as inflammatory-regenerative and 96 as dysplastfc (38 miid,
28 moderate and 30 severe dysplasia). On paraffin sectfons 3 monoclonal
antibodies against various leukocytic antigens were used.
Using a semiquantitive method the quantity of mononuklear inflammatory
cells was defined, their types, distribution, and altered interaction with
epithelial cells.
Results CD45R0 T lymphocytes were predominant cells in
the infiltrates. Less frequent were CD 68 macrophages. CD20 B lymphocytes
within the inflammatory cells was generally Iow.The intensity and composition
of the local immune reaction increased proportionally with the intensity
of epithelial dysplasia.The lymphoreticular cells were accumulated mainly
in the lamina propria. Direct contact between cells of the lymphoreticular
infiltrate and dysplastic epithelial cells was found rarely. Even where
close contact between lymphocytes and macrophages and dysplastic cells
was found, no signs of direct lytic effects on the dysplastic cells were
observed. It seems that dysplastic epithelial lesions do not provoke a
signifcant defence immune response, but the local immune reaction might
be a part of etiologically different bowel mucosa inflammation which may
lead to lesions.
Conclusion Our study proved that specific local immune
response in dysplastic epithelial lesions in flat bowel mucosa cannot be
used as important prognostic factor.
Pathology
and pathogenesis of African swine fever.The role of the monokines
Gomez-Villamandos J C,
Carrasco L, Salguero FJ, Mekonnen T, Ruiz-Villamor E, Bautista MJ, Sierra
MA
Dept. Anatomia Patologica
Facultad de Veterinaria Cordoba, Spain
African swine fever (ASF) is perhaps the most controversial and interesting
model of a swine disease with monocyte macrophage as the main target cell
for virus replication. The aim of this paper is to show the relationship,
in space and time, between macrophage alterations, including pro-inflammatory
monokines expression, with vascular lesions, lymphopenia and virus replication
in non-MPS cells. A pathogenic mechanism is proposed for the disease. Large
White x Landrace cross-bred pigs of both sexses were used for this study
The animals were inoculated, by the intramuscular route, with 105 50% HAD50
of the E70 ASF-virus isolate. Tissue samples were routinaly processed for
electron microscope. Immunohistochemical study to detect IL-Ia and TNF-a
expression was carried out.The pathogenesis of hemorrage is related to
the stimulating of endothelial cells and the subsequent loss of the endothelial
lining, virus replication takes place in these cells in the f nal stage
of the disease. Loss of endothelium allows platelets to adhere to the basement
membrane aggravating DIC, which commences prior to the appearance of this
lesion. ASF-infected swine suffers severe lymphopenia, that suddenly appears
during the middle phase of ASF Our studies have shown that the destruction
of lymphocytes in lymphoid organs and in the cellular infiltrate is due
to apoptosis. Immunohistochemical study revealed an increased level of
IL-Ia and TNF-a expression in these animals. Our finding constitutes the
experimental evidence underlying the hypothesis that monokines released
by activated macrophages and/or infected macrophages play a major role
in the pathogenesis of acute ASFThis study was supported by D.G.E.S. (Spain)
grant No. PB95- 158.
Pathological
studies on ovine caseous lymphadenitis in Turkey
Sayin Fatma
University of Adnan
Menderes, Faculty of Veterinary Medicine, Department of Pathology Turkey
Caseous lymphadenitis caused by Corynebacterium pseudotuberculosis
in small ruminants has been reported in many countries of the world (4),
includingTurkey For some years, this disease in sheep has become the most
important problem of Government farms in Turkey, by reducing both meat
and wool production, and by decreasing fertility (1,2,3).This study is
aimed to examine the pathology of this topic.The disease was inspected
in sheep, belonging to Ceylanpnar farm in southern part ofTurkey, in March
1997. Materials were taken from the lesional regions, and fxed in 10 %
neutral formalin solution. After paraffin-embedding, 5mm sections were
prepared and stained with hematoxylin- eosin for histological examination.
Lesions of caseous lymphadenitis were seen in 23 of 75 clinically suspected
animals. Grossly, the typical finding was the abscess formation. They were
seen as capsulated nodular lesions in different size and consistency The
small, new abscesses were of a pasty consistency while the large, older
abscesses were dry and solid.The color of necrotic abscess material was
greenish-yellow; there was no odor to them, and cut surFace of some showed
a concentric Iamellation.The abscesses in the lung were measured 1-4 cm
in diameter, whereas those in liver were 0.5-3 cm, and those in lymph nodes
were ranged from 0,5 cm to almost as large as the organ itself. Gross abscesses
were seen generally on mandibular (10 cases) and mediastinal (8 cases)
lymph nodes, followed by scapular, femoral and bronchial lymph node abscesses
in decreasing numbers. Lesions were only seen in lung (7 cases) and liver
(2 cases) from the visceral organs. Microscopically, the necrotic areas
in the center of the abscess consisted of destroyed neutrophylic leucocytes.
It was surrounded by a layer of mainly macrophages and some mononuclear
ceIIs.The capsule was made of fibrous tissues. Morphologic and immunohistochemical
studies are still going on.
Literature: 1. Batey, R.G.(1986).AustVet J. 63(4) : 131.
2. Paton, M.W et all. (1988).AustVet J. 65(4) : 117
3. Paton, M.VV et all. (1994). AustVet J. 7 1(2) : 47. 4. Pepin, M.
et all.(1993). Am J Vet Res. 54(6) : 873.
First
report of Peste des Petits Ruminants in lambs inTurkey: Patomorphological
and immunohistological feature
Alcigir, Gunay*; Yural
Atalay Sevil*; Toplu, Nihat**
*University of Ankara,
Faculty of Veterinary Medicine, Department of Pathology Ankara, Turkey
** University of Adnan
Menderes,
Faculty of Veterinary Medicine, Department of Pathology, Turkey
Three dead and one live animal were chosen from a flock consisting
of 6-7 month old lambs with 210 lambs from the district ofAnkara in 1993
for the determination of an illness showing symptoms of fever, cough, diarrhea
and had 30 % mortality. By using pathologic and immunhistologic methods
the disease was diagnosed as Peste des Petits Ruminants. Grossly, light
necrotic and erosive stomatitis and few erosions in abomasum were identified,
1-2 mm diameter white-gray foci in liver, hyperemia and edema in intestines
were seen. In cranial and caudal lobes of lungs there were 2-5 cm diameter
red sometimes gray hepatization areas. Histopathological signs were characteristic
and there were pathognomic multinuclear syncytial giant cells and intracytoplasmic
inclusion bodies in mucosa of tongue and in abomasal glandular cells. Syncytial
giant cells with few nuclei were detected in mediastinal and mesenterial
lymph nodes. Although there was small and irregular focal necrosis in liver,
the main sign was the finding of syncytial giant cells with 3-5 or more
nuclei and some intranuclear inclusion bodies. The most evident signs in
diagnosis of the disease were observed in lungs. Mainly puruVent, sometimes
desquamative catarrhal bronchopneumonia and necrotic areas were apparent.
Interalveoler intersititiel pneumonia was present between these areas.
Some bronchial epithelial cells were hyperplastic. In addition to these
characteristic signs, the diagnosis was supported by the detection of multinuclear
syncytial giant cells in alveoler lumina and intracytoplasmic inclusion
bodies in bronchi and bronchial glandular cells and sometimes pneumocytes.
On the other hand, reddish stained diffuse or granular viral antigens were
detected by Avidin-Biotin Complex (ABC) method using rabbit anti-rinderpest
hyperimmune serax within the lungs in bronchi and bronchioler epithelial
cells, bronchial glandular cells, oral mucosal cells, hepatocyts, particularly
reticulum and macrophage cells in lymph nodes. The macroscopic and microscopic
structures of the lesions were in accordance with the literature data (1,2,3),
except the finding of liver lesions.
Swine
leptospirosis caused by Leptospira interrogans (serologic variant Icterohaemorrhagiae)
Modrić Z, Sabočanec R,
Turk N, Artuković B, Slunjski A, Čuljak K
Department of General
Pathology and Pathological Morphology
Faculty of Veterinary
Medicine of the University of Zagreb, Zagreb, Croatia
In
one of nine piggeries of one pig herd, 18 days after sows farrowed litters
some of piglets were severely jaundiced and poor viable were died. Five
of this sick piglets had titres for leptospires but twelve sows ofthem
had not.Total of fme piglets which were brought to our clinic due to determine
diagnosis, had aglutinins for leptospires and pathological changes in one
died and two sacrified piglets were severe jaundice, liver and kidney parenchymatous
degeneration and splenomegaly. From sacrified piglets kidneys a leptospires
serogroup icterohaemorrhagiae were isolated by cultivation in Korthof's
media.
In two piglets a streptomycin was administered during four days what
was resulted with failure in isolation of leptospires from theirs kidneys.
The obtained facts indicates that the source of infection for piglets
in piggeries were very probably rats.
The
comparison of histopathological, immunoperoxidase and immunofluorescence
techniques in the diagnosis of rabies in dogs
Vural Atalay Sevil
University of Ankara,
Faculty of Yeterinary Medicine, Department of Pathology Ankara, Turkey
The effectiveness of the immunoperoxidase staining method for the diagnosis
of rabies was investigated in this study, compared to immunofluorescence
and histopathological methods.The tissues (cornu ammonis, cerebellum, thalamus,
colliculus rostralis, pons cerebri, nucleus caudatus, medulla oblangata,
spinal cord, Gasserian ganglion, parotid gland, submandibular salivary
gland, adrenal gland, retina, cornea, trachea, stomach, intestine, mouth,
nose, ear, and urinary bladder) of 10 rabies dogs were examined.The samples
were fixed in 10 % bufferformaline and were blocked in paraffin and sectioned
at 4-5 mm.All sections stained with hematoxylin and eosin and Avidin-Biotin
Complex (ABC) and indirect immunoflourescent test using rabbit anti-rabies
hyperimmune sera*.The nonspecific inflammatory reactions and degenerative
changes detected by histopathologic methods regarding rabies were observed
in the central nervous system, Gasserian ganglion, parotid gland, adrenal
gland, and submandibular salivary gland.The Negri bodies were seen in the
cornu ammonis, cerebellum, thalamus, nucleus caudatus, colliculus rostralis,
and also, in the Gasserian ganglion. The immunoperoxidase method, showed
extra and intra cellular rabies virus antigen and/orthe inclusion bodies
in the entire central nervous system and Gasserian ganglion. In addition,
immunopositive rabies virus antigens were also identified in retina, submandibular
salivary gland, cornea, parotid gland, trachea, stomach, intestine, ear,
mouth, nose, urinary bladder, and adrenal gland. The immunofluorescence
method, revealed rabies virus antigens in the central nervous system, Gasserian
ganglion, retina, parotid gland, ear and mouth. In this study, equally
positive results were obtained from the avidin-biotin peroxidase and indirect
immunofluorescence method. Although both methods could be used conveniently
in formalin fixed and paraffin embedded blocks, the immunoperoxidase method
appeared to be more advantageous for use in pathology laboratories since
this method permitted usage of high titered sera as well as usage of more
simplified microscopes and storage of stained slides for a long time. This
immunoperoxidase method has been used forthe first time forthe diagnosis
of rabies in our country.The microscopic structures of the lesions were
in accordance with the literature data (1,2,3,4).
Literature: 1. Balachandran,A. et all. (1994).Vet Pathol. 31:
93.
2. Das, S.K. et all. (1985). J Anim Sci. 55 : 979.
3. Goldwasser, R.A. et all. (1958). Proc Soc Exp Biol Med. 98 : 219.
4. Sinchaisri,T et all. (1992). J Vet Med Sci. 54 : 409. *Provided
Centre of Expertise for Rabies Agriculture Canada Animal Disease Research
Institute Nepean, Ontario.
1 Brown, C.C et all. (1994).Vet Pathol. 31: 194
2. Brown, C.C et all. (199 1).Vet Pathol. 28 : 166.
3. Obi,TU. et all. (1983). ZbI Vet Med B. 30 : 751. Provided Animal
Disease Research and Control Institute, Ankara,Turkey.
Implications
of pulmonary intravascular macrophages in pulmonary processes
Carrasco L, Gomez-Villamandos
JC, Bautista MJ, Sanchez-Mascaraque C, Salguero FJ, Millan Y, Quezada M
Dpto Anatomia y Anatomia
Patologica Comparadas, Cordoba, Spain
In recent years the study of the pulmonary physiology and the pathogeny
of pneumonic process of certain species of mammals has suffered important
changes, due to the description of a resident population of cells to the
Mononuclear Phagocyte System, the pulmonary intravascular macrophages (PIMs),
which form a resident population in the pulmonary capillaries of certain
animals. The existence of PIMs was first reported in the early 70's, but
they were not studied in any great depth until the late 80's, when several
studies of their features and functional significance were made in goat,
bovine, sheep, deer, swine, horse and rabitts.There is evidence that, like
macrophage from other sources, PIMs have ample secretory capabilities and
influence the pulmonary microvascular physiology through release of proinflamatory
mediators that can initiate or enhance acute pulmonary inflamation. However
there are few studies that have studied the implications of the PIMs in
the development of the acute process that characterized some diseases.
From this reason we have analyzed the implications of PIMs in animals experimentaly
infected with some viruses, such as African Swine Fever, African Horse
Sickness, Rabbit Haemorrhagic Disease; and bacterias as Pasteurella haemolytica
and characterized by the establishment of an acute pulmonary process. In
these infections we have observed that PIMs sufferer and activation processes,
from the first time of infection, were associated with different pulmonary
pathological changes, such as neutrof I accumulation in septal capillaries,
pulmonary endothelial injury, increased microvascular permeability with
a consequent interstitial and alveolar edema and microvascular thrombosis;
in African Horse Sckiness and Pastereullosis a concomitant pneumonic proccess
was found, characterized by the existence of alveolar macrophages activation
and fbrin deposits in alveoli.We can conclude that although the existence
of PIMs should be benefical to the animal, since they actively participate
in blood clearance, interactions with different antigens, subsequent activation
is, also, associated with pulmonary pathological changes.
Incidence
and pathological studies on the liver telangiectasis on feedlot cattle
Atasever, Ayhan*; Vural
Atalay, Sevil**; Berkin, Senay**
University of Erciyes,
Faculty of Veterinary Medicine, Department of Pathology Turkey ** University
of Ankara, Faculty of Veterinary Medicine, Department of Pathology, Ankara,
Turkey
The livertelangiectasis is a fairly common lesion offeedlot cattle
which causes important economic losses (1).This report deals with the gross
and microscopic pathology and incidence of telangiectasis. In this study
3470 feedlot cattle slaughtered in Ankara Meat and Fish Company abbatoir
were inspected. Relationship of abscesses to telangiectasis was also examined.
All of the tissue samples were collected in a few hours following the slaughter.The
samples were fxed in 10% buffer formaline and were blocked in paraffin.All
the samples were stained with hematoxylin and eosin, Best, Luna,Toluidine
blue, van Gieson and Gram stains.Telangiectasis was observed in 82 of 3470
livers (2.1%). Lesions were mostly localized on the parietal surface of
the lobus dexter, and occasionally generalized throughout the organ.They
were seen grossly as single or multiple (in 2 cases) reddish-brown foci.
Each focus was 1to 3 mm in diameter, occasionally larger. Microscopically,
they were generally characterized by distortion and necrosis of hepatocytes
with dilatation and congestion in sinusoids.The lesions were observed in
74 livers with normal appearances. Multiple abscesses were observed in
6 livers formed far away from the telangiectasic lesions. Lesions of severe
distomatosis were observed in 2 cases. The macroscopic and microscopic
structures of the lesions were in accordance with the literature data (1,2,3).
No correlation was found between telangiectasis lesions and liver abscesses.
This is the first evaluation of this subject in Turkey.
Literature: 1. Getty, R. (1946).Am J Vet Res. 7:437
2. Jensen, R. et all.(1982).Am J Vet Res. 43 : 1436
3. Jensen, R. et all.(1947). J A V M A. 110:256.
The
differences in pathological picture in pigs succumbed of hog cholera
Šoštarić B, Lipej Z
Croatian Veterinary
Institute, Zagreb, Croatia
Hog cholera recently appeared as a major threat to the European pig
industry, whereas it has been present in Croatia, as an endemic disease,
during a long period of time. During several decades systematic diagnostic
proceeding has been carried out at the Department of Pathology of the CroatianVeterinary
Institute; and during that time thousands of animals succumbed due to hog
cholera were dissected and pathoanatomicaly evaluated. It has been noted
that the pathoanatomical picture had changed during that period, and that
lesions earlier considered as pathognomonic are not a useful diagnostic
value any more, even more, some of them are not present as a rule, or they
are rearly found. Recent pathoanatomical fmdings are more specifically
described in this paper.
Immunohistochemical
identification of Leptospira pomona antigen in tissue sections
Juntes P, Lindtner R,
Pogačnik M
University of Ljubljana,
Veterinary Faculty, Ljubljana, Slovenia
Aim Post-mortem diagnostic of leptospirosis in domestic animals
is a difficult task. One way how to do it is by using immunohistochemical
procedures (1,2,3). The aim of our study was to produce polyclonal antibodies
in rabbits which could be used for immunohistochemical identification of
Leptospira interrogans serovar pomona in routinely prepared paraffin embedded
tissue sections.
Methods Rabbits were immunised with non-pathogenic serovars
of Leptospira pomona using partly modified previously described procedure
(3).The culture for immunisation contained 10 8 Microorganisms and was
administered intravenously three times.Animals were bled one week after
last application. Sera were tested on smears of Leptospira pomona culture
and on formalin fxed paraff n embedded tissue of several animal species
DAKO En Vision +(tm) System, was used for the visualization of immunohistochemical
reaction, with 3-amino-9-ethycarbazole (AEC) as a substrate. Positive antiserum
against Leptospira pomona and positive tissue (a gift from dr. Eugenio
Scanziani, Italy) were used as a positive control and incubation without
specific antibodies as a negative control.
Results and conclusions Antisera that we produced gave
a strong and clear immunohistochemical reaction on paraffm tissue sections
fixed in buffered formalin.With a very sensitive kit for visualization
that we used, we were able to detect leptospiral microorganisms even on
material that was not properly fixed. Pattern of reaction products produced
on culture smears was granular without recognizable leptospiral morphology.
It was confirmed that immunostaining is useful for the detection of Leptospira
pomona infections in animal tissues.This procedure is especially valuable
in cases when microorganisms in tissue are not viable or microbiological
isolation is not possible for other reasons.
References 1) EllisTM et al.AustrVet J I983;60:364-367;
2) Poonacha KB et aLVet Pathol I993; 30:362-369;
3) Scanziani E. ResVet Sci 1991; 50:229-232.
Pathomorphology
of vibriosis in sea bass (Dicentrarchus labrax) of different ages
Zrnčić S, Oraić D, Šoštarić
B
Croatian Veterinary
Institute, Zagreb , Croatia
Vibriosis caused by Listonella (Vibrio) anguillarum is one ofthe most
devastating diseases in cage cultivated sea bass (Dicentrarchus labrax)
in Croatia.This work derives from our observation of different fish farms
alongthe coast systematically monitored forfish diseases. Pathomorphology
of this bacterial disease in different ages of sea bass is presented in
this paper.The affected fish in all observed cases swam slowly to the water
surFace, separated from the lot and were reluctant to eat. Pathoanatomically,
fish showed skin decoloration, presence of skin erythema on the f n base,
around the mouth, on the vent and in more advanced cases ulcerations were
noted as well. Internal examination revealed liver, stomach, intestine,
fat tissue, gonads and swim bladder hemorrhages and enlarged spleen.The
samples of tissue of all macroscopica!!y changed organs were submitted
to histopathological examination and the results are presented in this
paper.
Immunopathological
response against Trichinella spiralis in mice
Marinculić A*, Božić
F*, Dominis M**, Auslender Ujević V**, Grabarević Ž*, Duraković E*
*Veterinary Faculty
** Pliva
*** Veterinary Institute,
Zagreb
By the first quantitative phenotypic analysis
of intestinal intraepithelial lymphocytes (i-IEL) at the early stages of
theTspiralis infection, we found that the kinetic of ((T cells in i-IEL
corresponded to the kinetics of worm expulsion after infection of C57BL
mice with 200 muscle larvae. Infected mice developed signifcant enteropathy,
comprising villus atrophy, crypt hyperplasia, goblet cell hyperplasia and
a decrease in i-IEL numbers by 14 days post infection (p.i.), when most
of the parasites had been expelled. Most murine i-IEL of the gdT
cell lineage tend to be cytolytic when activated.We speculated that gdT
cells of i-IEL during the early stages of infection recognize and eliminate
damaged epithelial cells generated by parasite antigens, simultaneously
accelerating the worm expulsion. However, since the rejection ofT.spiralis
worms is dose-dependent effect, to assess whether or not i-IEL, particularly
gdT
cells, respond comparably to different sized T.spiralis infection, we used
C57BL mice, orally infected with 50 or 400 viable T.spiralis larvae. We
found that the rapid activation of gdT
cells in i-IEL occured much sooner in mice infected with 400 larvae than
in mice infected with 50 worms, as early as 1 day p.i. vs 4 day p.i., respectively
In addition we found significant elevations of eosinophiles and mastocytes
on day 1 p.i. in mice infected with 400 T.spiralis larvae in comparision
with mice infected with 50 larvae. In spite their intrinsic cytolytic activity
against transformed cells, no corelation between kinetics of gdBT
cells in i-IEL and T.spiralis worm expulsion was observed in mice infected
with 400 or 50 Iarvae.These fndings strongly suggest that bothTspiralis
infection levels elicits immunodepression by interFerence with the first
line defenceT cell function, therefore influence the outcome of experimental
T.spiralis gut infection.
B.
Iatrogenic,
experimental and environmental pathology
Iatrogena,
eksperimentalna i patologija okoliša
The influence
of thyroid functional modifications on collagen contens of skeletal muscle
in the meat chickens
Curca Dumitru
Department of Pathophysiology,
Faculty of Veterinary Medicine, Bucharest, Romania
The collagen contents ofthe adult animal
organisms is generally constant, but in some physiological states (parturition)
and especially in physiopathological ones, marked modifications take place,
both quantitatively and qualitatively. Endocrine glands through the hormones
they produce, influence both the quantity and the quality of collagen,
thyreoptropine is a stimulator mainly for the hyaluronic acid in the ground
substance, while thyroxine stimulates the connective tissue in thyroidectomized
animals. Experimental research was conducted on three batches of broilers
aged 40 days. Batch I. consisting of eight broilers having a weight of
837.50+/- 13.98 g to be brought into a condition of hyperthyroidism, was
administred 2 mgTIROTON/ 5kg of feed, recipe 2 1/1in their combined feed
that is about 40mg
TIROTON per broiler each day. Batch II. consisting of eight broilers having
a weight of 677.50 +/- 20.68 g to be brought into a condition of hypothyroidism,
was administred 5 g METILTIOURACIUS kg offeed in their combined feed,which
means 100 mg METILTIOURACIL per broiler daily. Batch III was the control.
After 42 days of feeding, the broilers were slaughtered with a view to
dosing the collagen in their muscular system by means of the indirect biochemical
metod which aims at the determination of the hydrowyproline contents according
to Neuman and Logans techical indications. Broilers whose feed was supplemented
withTIROTON showed a statistically insignificant decrease tendency of collagen
in the pectoral muscle (P0.05), while broilers in the batch treated with
METILTIOURACIL showed a statistically insignificant increase tendency (P0.05).
In the thigh muscles, both batches showed an increase tendency, but statistically
insignificant (P0.05).The body weight in both batches was smaller (P<0.01
) as compared to the controLThe histopathologycal examination ofthe thyroid
in the broilers treated withTIROTON revealed the flattening of follicles
that are full of colloid. In the broilers treated with METILTIOURACIL,
the thyroid follicles are unequal, some of them quite large, the walls
become thickened, the colloid sometimes having a haemorrhagic aspect owing
to their cystization, which characterises the condition of hypothyroidism.
Pathomorphological
picture of Myxidium infection in cultivated charp snouted sparus (Puntazzo
puntazzo)
Oraić D, Zrnčić S, Šoštarić
B
Croatian Veterinary Institute,
Zagreb, Croatia
Charp snouted sparus (Puntazzo puntazzo) is a native inhabitant of
the Adriatic sea.This species has not been cultivated in our fish farms
until recently.At a certain fish farm, experimental production of charp
snouted sparus started during 1997., using imported fry. Seven weeks old
fry was put into growing cages, but after two months high mortalities started.
At the beginning, affected fsh swam slowly close to the surface separated
from the lot and they were emaciated. Pathoanatomically only slight patchy
skin discoloration was observed. By internal examination of recently dead
and euthanized moribund fish stomach appeared normal, and was filled with
usual amount of ingesta, while immediate proximal anatomical structure
of intestine was distended by seromucous transparent content.The intestinal
wall was thinner and more translucent then normal. Examination of gut mucosa
scrapings using light microscopy (400x) disclosed numerous spores of myxosporean
parasite Myxidium leei. Histopathological examination of intestines revealed
numerous parasitic structures as well, and therefore confirmed the diagnosis
of M. leii.
Use
of LN3 antibody in the analysis of the biological evolution of myocarditis
Bassan F, Bergamin C,
Bussani R, Pavletic N, Silvestri F
Institute of Pathological
Anatomy - University of Trieste, Italy
Myocarditis can def nitively be diagnosed only by endomyocardial biopsy.
The traditional histology may be used to: 1) diagnose a "Dallas" myocarditis
2) analyse the myocarditis evolution 3)? evaluate the type and grade of
endomyocardial fibrosis 4)? evaluate the myocytes (hypertrophy; atrophy;
loss of contractile material, etc.) 5) evaluate the aetiology of some infective
myocarditis (bacterial, fungal, protozoal, etc.) What is actually needed
is not only an accurate histopathological diagnosis of myocarditis, but
also to find some criteria that could be used for identifying the "non
Dallas" myocarditis and some prognostic determinants for the patients affected.
To assess the overall status of"immunological activation" of the heart
we think that the antisera that perform correct immunophenotypical characterisation
of inflammatory cardiac infiltrates are: CD45R0, CD45RA, CD4, CD8, CD20,
LN3, and CD68-PGM-I. Monoclonal antibodies directed against T or B-cell
antigens facilitates identification and quantifcation of inflammatory infiltrates.
LN3 (HLA DR-expressing cells) seems to be a particularly useful marker
for detection of a generalised autoimmunological process affecting the
myocardium. Endomyocardial biopsy (EMB) has been perFormed, at our Cardiology
department, on eight-hundred thirty patients with suspected myocardial
disease of undef ned aetiology from 1978 to 1996. In 60 patients an active
myocarditis has been identifed. The first biopsy showed HLA-DR molecules
in all these patients. In the follow-up biopsies LN3 expression in lymphocytes,
myocytes and in endothelial cells was evident (43%) only in patients with
ongoing myocarditis or in those with dilated cardiomyopathy (DCMP) evolution.
A retrospective study evidenced, moreover, LN3 expression in EMB of 10
patients with clinical diagnosis of DCMP and traditional histology positive
for"borderline" myocarditis. LN3 may be a diagnostic tool for identification
of the "non Dallas" myocarditis and a useful marker in the evaluation of
prognostic determinants for patients with myocarditis.
Use
of microwawe in a histopathology laboratory
Bassan F, Zambon S, Bussani
R, Pavletic N, Silvestri F
Institute of Pathological
Anatomy - University of Trieste, Italy
Microwaves are very much in use for performing special staining and
are also being used for other procedures in Histotechnology. The microwave
electrical field that is produced creates friction as it passes through
dipolar molecules causing them to reverse themselves 2450 million time/sec.
The use of the microwave was firstly introduced in immunohistochemistry
applications, but only recently its use extended to the field of histochemistry.
Microwaves have been a great property of reinstating immunogenetic cells
to sever every cross-linking formed by fixative and proteins (NH2 -COOH).
Histochemistry is able to reduce greatly the time of tissue incubation
in staining procedures and especially when silver-nitrate impregnation-method
is concerned. We found particulatly staining techniques for polysaccharides,
neutral mucosubstances and basement membranes demonstration using PAS reaction,
for connective and muscle tissue and glial fbres with Azan Mallory techniques.
The best result that we got in this field has been the silver-nitrate impregnation
for Borrealiae checking in paraffine-embedded tissue sections.We elaborate
a special method that clearly underlines Spirochaetes reducing to a minimum
the background staining.
Effect
of colhicine on redistribution of cell membrane proteins in the rat proximal
tubule
Baus M, Medjugorac-Popovski
M, Sabolić I
Institute for Medical
Research and Occupational Health, Zagreb, Croatia
Epithelial cells along the mammalian nephron and in other tissues establish,
maintain, and modify their plasma membrane composition by the process of
membrane recycling (Brown D and Stow JL, Physiol. Rev. 1996, 76:245). In
general,the recycling of various membrane components between the plasma
membrane and intracellular organelles by means of endo- and exocytosis
depends on intact microtubules (MT). However, the details of the recycling
mechanism for specific plasma membrane proteins are poorly understood.ln
the renal proximal tubule (PT) cells,the rate ofthe MTdependent recycling
of a particular protein may be related to its localization in the specifc
plasma membrane domain (brush-bordervs. basolateral) orto its association
with the membrane, e.g. whether a protein is an endo-,transmembrane-, or
ecto-protein. Another discriminating factor in this recycling may be the
role the specific protein plays in acute or chronic adaptations to the
tubular load with the filtered substrate. In this work, we used an MT depolimerizing
agent, colchicine, to study the distribution of Na/K-ATPase [a basolateral
(BLM) transmembrane protein], vacuolar proton-ATPase [V ATPase; a brush-border
membrane (BBM) endo-protein], sodium-phosphate co-transportertype 2 (NaPi-2;
a BBM transmembrane protein), and cell adhesion molecule CAM-105 (a BBM
ecto-protein) in the rat kidney PT cells. Rats were injected i.p. with
a single dose of colchicine (0.35 mg/ 100 g B.M.) or phosphate-buffered
saline (controls).Twelve hours later, the animals were anesthetized and
kidneys were fixed by perfusion in vivo. An indirect immunofluorescence
cytochemistry in 4 (m thick cryosections of the kidney cortex was used
to label specific membrane proteins in the PT cell plasma membrane domains
and intracellular vesicles. A pattern of staining with the monoclonal and
polyclonal antibodies to the specifc membrane proteins, followed by the
fluorescein- or CY3-labeled secondary antibodies, showed in colchicine-treated
rats: a) disappearance of MT in the PT cells, b) marked loss of the 31
kDaV ATPase sub-unit and NaPi-2 from the BBM, and their redistribution
in numerous membrane vesicles randomly scattered in the cell cytoplasm,
and c) unchanged distribution of the CAM- 105 and Na/K-ATPase in the BBM
and BLM, respectively. We conclude that the abundance of V ATPase and NaPi-2
in the BBM, e.g. transporters whose concentration in the PT cell membrane
is known to adapt quickly to acute changes in acid and phosphate loads
in the PT, respectively, are very sensitive to MT depolimerization.The
slowly-recycling membrane proteins, such as the Na/ K-ATPase in the BLM,
and the CAM- 105 in the BBM, are not significantly affected by MT disruption.
Serum cholinesterase
activity (PchE) and ascorbinemia calves with neonatal diarrhoea after parenteral
administration of ascorbic acid
Curca Dumitru
Department of Pathophysiology,
Faculty of Veterinary Medicine, Bucharest Romania
Comprehensive studies made on ascorbic acid's (AA) multiple biochemical
function have shown that the demand for this vitamin, to get healthy calves
with optimum perFormances, is signifcantly higher than the quantity needed
to prevent some clinical deficiencies, being noticed an obvious difference
between the minimum and optimun needs. Synthesis and using of AA is feed-back
regulated, being influenced by living conditions as food, climate, biologic
agents action, to which the calves functional peculiarities, like the hormonal
and metabolic characteristics are added. A study was made of the total
blood level of AA by the Roc and Kuether method, and of PChE activity by
the Ellman et al. method, in calves with moderate diarrhoea, after intravenous
administration of AA 1g/day for 4 successive days, as compared with untreated
calves and with clinically healthy congeners.A statistically significant
decrease of ascorbinemia (P<0.05) and PChE activity (p<0.0 1) was
found in calves with moderate diarrhoea aged 4 to 14 days as compared to
congeners clinically healthy calves, whereas the hematocrit did not show
statistically signifcant modifications (P0.05). After the daily intravenous
administration of 1g AA for 4 successive days an increase of ascorbinemia
to 3.289 +/- 0,533 mg/dl blood was noted in calves with neonatal diarrhoea,the
difference being statistically nonsignificant (P0.05) versus nontreated
clinically healthy calves. After treatment, the ascorbinemia level was
restored, but the PChE activity remained low, 176.475 +/- f 6.01 mU/ml
blood serum,the difference being neatly significant (P<0.01) as compared
to nontreated clinically healthy calves. Nevertheless, there was an improvement
of appetite, a restoration of the habitus, although no correlation was
noted with the increase of the PChE activity underthe conditions of statistically
nonsignifcant (P0.05) modifications ofthe hematocrit. The results also
reveal the existence of a relatively wide variation of AA and PChE activity
in calves, depending on age, state of health and parenteral administration
of AA.
Polyclonal antibodies
to rat renal cortical endocytic vesicles labe4 proximat tubule cell rnembranes
In the rat and human kidney
Međugorac-Popovski M, Herak-Kramberger
CM, Sabolić I
Institute for Medical Research
and Occupational Health, Zagreb, Croatia
Proteins that are f Itered in the renal proximal tubule (PT) are reabsorbed
by endocytosis, a process which occurs via specialized intracellular organelles
- endocytic vesicles (EV, endosomes). Due to imperFect methods, EV are
hard to isolate without a significant contamination with other (intra)
cellular membranes.The aim of this study was to produce specific polyclonal
antibodies to the antigens in endosomal preparations that could be used
either in immunoisolation experiments to get cleaner preparations of EV
and other organelles or/and for specific labeling of the cell plasma membrane
(PM) domains and intracellular structures in animal and human kidney. Endosomes
were isolated from the rat kidney cortex homogenate by the established
method (Sabolia I. and Burckhardt G., Methods Enzymol. 1990, 191:505),
and characterized functionally and enzymatically. The final vesicle preparations
exhibited high activity of the baf lomycin-sensitive proton-pump (V ATPase)
and minimal contamination with the brush-border (BBM) and basolateral membranes
(BLM). (mmune sera to the EV preparations were raised in rabbits. As found
by indirect immunofluorescence cytochemistry in 4 (m thick cryosections
of the rat kidney cortex, the immune sera brightly stained the BBM and
various intracellular organelles in the PT cells. In immunoblotting experiments,the
sera strongly labeled 4 high (Mr 55-276 kDa) and 5 low molecular mass (16-36
kDa) protein bands. However, none of the bands were specific for EV.An
antibody to the 104 kDa protein band was purifed by immunoadsorption. This
antibody strongly stained the BBM and subapical vesicles in PT cells of
the rat and human kidney cortex. In the human kidney cortex, it also stained
the BLM of some collecting duct ceIIs.Thus, common antigenic domains exist
in the cell membrane proteins in rat and human kidney tubules. Conclusion:
Polyclonal antibodies to the rat renal EV failed to specifically label
endosomes and, therefore, could not be used for better purification of
these organelles. The antibodies, however, might be used in cell biology
studies of the common membrane antigens along the rat and human nephron.
Nephrotoxic heavy
metals increase the expression of mdrl P-glycoprotein in the rat kidney
brush-border membrane
Herak-Kramberger CM, Thevenod F*,
Sabolić I
Institute for Medical Research
and Occupational Health, Zagreb, Croatia
*II Physiologisches Institut,
University of Saarland, Homburg/Saar, Germany
The multidrug resistance P-glycoprotein (mdr 1) is an 150- 170 kDa
membrane protein that catalyzes the ATP-dependent efflux of hydrophobic
and potentially cytotoxic compounds from the cells and, thus, it may represent
an important cellular mechanism of detoxication. In various epithelia,
including the kidney proximal tubule (PT), the protein is expressed on
the apical cell membrane. An altered expression of mdrl is commonly seen
in tumors treated with chemotherapy or in some cultured cells exposed to
environmental stress or cytotoxic heavy metals (1). Using a monoclonal
anti-mdrl antibody (C2 19), we studied the expression of mdr 1protein in
PT cells by indirect immunofluorescence in 4 (m thick frozen sections of
the fixed kidney cortex and by immunoblotting in isolated renal cortical
brush-border membranes (BBM) from control rats and from the rats treated
in vivo with various heavy metals. Rats were treated with daily s.c. injections
of chloride salts of Cd, Hg, Cu, Zn, Mn, Ca, Mg, AI, La (2 mg/kg B.M.)
or Pb (5 mg/kg B.M.) for 14 days, or with a single i.p. injection of cis-Pt
(5 mg/kg B.M. 5 days before sacrifce). Control rats received an equivalent
amount of saline for a comparable time period.A relative amount of mdrl
in BBM was correlated to the metal concentration in the renal cortical
tissue measured by atomic absorption spectrometry. ln control rats,avery
lowtissue concentration ofthe metals was detected and a heterogeneous expression
of mdr 1in BBM along the PT was found (S 1<S2<S3). In metal-treated
rats, the concentration of metals in the renal cortical tissue increased
between 7-(Mn) and 3000-fold (Cd). In rats treated with Cu, Zn, Mn, Ca,
Mg,AI, and La, the abundance of mdr 1in the cortical BBM was not different
from that in controls. On the contrary, in rats treated with the well-known
nephrotoxic heavy metals, such as cis-Pt, Pb, Cd, and Hg, the mdr 1expression
in the cortical BBM increased 3-, 6-, 15-, and 18-fold, respectively. Conclusion:
nephrotoxic heavy metals (Cd, Hg, Pb and cis-Pt) increase the expression
of mdrl in the rat kidney cortex BBM in vivo.The increased expression of
mdr 1may represent a part of a specific cytoprotective response of the
PT cells to oxidative stress induced by nephrotoxic heavy metals. Functioning
as a lipid-translocase (2), mdrl could assist in removal of toxic lipid
peroxides generated in the presence of heavy metals by translocating them
across the apical membrane into the PT lumen.
References: 1. Gottesman MM and Pastan I, Annu. Rev. Biochem. 1993,
62:385
2.Van Helvoort A et al., Cell 1996, 87:507
Clinical and pathofogical
study in canine visceral leishmaniasis: natural and experimental infection
Marinculić A, Kamadan P, Petrinović
T, Grabarević Ž, Živičnjak T, Ćurić S, Stojčević D, Džakula N, Beck R,
Barbić A, Šantek T, Vukonić L
Veterinary Faculty, Zagreb,
Croatia
Canine leishmaniasis is a severe, often fatal, systemic disease of
the dog, caused by the protozoan parasite Leishmania infantum. The disease
is endemic in Mediterranean countries including Southern Croatia. It is
well known that the dog, apart from suffering the disease itself, is the
principal reservoir of the disease. In the dog as in other hosts leishmaniads
multiply inside macrophages causing chronic inflammatory processes .The
purpose ofthe study reported here was to compare the fndings of the clinical
aspects of the disease and the clinical pathology in naturally and experimentally
infected dogs with a field strain of Leishmania infantum. The special
aim was also to define the virulence and pathogenicity of the same strain
in dogs. In order to def ne the clinical aspect of the natural infection,
three dogs were hospitalized and thoroughly examined during the period
of 1month or up to the death. Among naturally infected dogs, different
dermatologic patterns were found like symetric alopecia, scaling and ulcerative
dermatosis beside cachexia and parakeratosis. According to the experimental
design, a group of six dogs was inoculated with parasites and the infection
lasted for 120 days. Samples were obtained from dogs prior to infection
and at weekly intervals for complete blood count and serum chemical analyses.
Significant changes in rectal temperatures and body weights were not observed
between infected and control animals over a 12 week period. All dogs developed
a persistent infection in the period of eight weeks The systemic infection
was defined by the parasitological findings of parasites in lymph node
biopsies. Infected dogs, also developed signifcantly altered hematological
values consisting of mild anemia and moderate leukopenia. Beside mentioned,
other clinical signs were not found until the end of the experimental period.The
lack of full-blown (eishmaniasis in the infected dogs is probably related
to the chronic nature of the disease in which long incubation periods are
ratherthe rule. It is not clearwhy in some dogs, clinical signs ofthe disease
are poorly manifested and in other dogs, the disease is fatal.
C.
"Herman Jurak"
round table on rheumathological diseases
"Herman Jurak"-
okrugli stol o reumatološkim bolestima
Anatomy
and biomechanical characteristic of joint
Nikolić V
Department of Anatomy,
Medical Faculty Zagreb, Croatia
Diarthroses or synovial joints are non continuous and
cavitated articulations with a fine capillary cleft between articular surfaces
(synovial cavity) which contains synovial fluid - synovia.The synovial
fluid is produced by the synovial membrane that lines the fibrous membrane
and whole articular cavity other than articular surfaces of the joint.
Bones are linked by the fibrous capsule and covered by articular cartilage.
The synovial membrane
The synovial membrane lines nonarticular surfaces in
synovial joints (fibrous capsules, intracapsular nonarticural bony surfaces,
intracapsular ligaments and tendons), bursae and tendon sheaths were movements
occur between contacting surfaces.The synovial membrane ceases at the articular
cartilage margin. Macroscopically it is smooth, pink and shining with small
synovial villi, folds and fringes on the internal surface. In some joints
accumulation of adipose tissue in the synovial membrane occurs in the form
of articular fat pads.The synovial membrane varies in local structure.
It has an interlacing discontinuous cellular layer with A and B -type synoviocytes
one to three cells deep, embedded in thin amorphous fibre free intercellular
matrix.The functions of the cells are: 1. Removal of debris from articular
cavity (mainly by type A cells). 2. Synthesis of the hyaluronic acid and
glycosaminoglycans, or proteoglycan - like material lubricin (which acts
as a lubricant of the articular cartilage) of the synovial fluid, and some
of the constituents of the extracellular matrix . 3. Some synoviocytes
produce antigens. Migrating synoviocytes carry ingested material to the
lymphatics of the synovial subintima. Division rate of synoviocytes increases
in response to acute trauma or haemarthrosis.
The synovial fluid
Occupies and can be found in synovial joints, bursae
and tendon vaginas. It is clear, viscous and slightly alkaline with a small
population of mixed ceIIs.Volume of synovial fluid in normal joints is
Iow.The composition of synovial fluid is similar to a dialysate of blood
plasma containing: proteins about 0.9 mg/ 100 ml, mucin, mostly hyaluronate
and glycosaminoglycan. Proteins are partly free and partly bound to glycosamonoglycans
and hyaluronate, 10 to 30% derive from blood plasma. The origins of some
other proteins are probably B type cells of synoviaf intima. A healthy
human joint contains approximately 1to 5 ml of synovial fluid.
Functions of synovial fluid are:
1.To provide a liquid environment forjoint surfaces,
internal fluid compartment and hydrodynamics of cartifage and menisci.
2. Nutrition of articular cartilage cells, discs and
menisci, acting as a medium for osmosis between the joint and capillary
network.
3. Lubrication and reductions of shear and erosion.
Synovial fluid has non-Newtonian properties, i. e.The
fluid is highly viscous with low rates of shear; with increased rates of
shearthe viscosity nonlinearly decreases, meaning that in slow movements
weight bearing capacity would be maximal. The product of viscosity and
shear rate is almost constant, and also the weight bearing capacity. Synovial
fluid obtained from degenerative joints show reduced non-Newtonian flow
properties compared to fluid from healthy joints.
Articular cartilage
The articulating bone surfaces are covered with 1to 5
mm thick articular cartilage. Articular cartilage's primary functions are
to distribute joint loads over a wide area decreasing the stresses on the
contacting joint surfaces and to allow movement with minimal friction and
wear.
Composition and structure of articular cartilage:
Articular cartilage is a special variety of hyaline cartilage
and consists of two compartments: cellular (chondrocytes) and extracellular
(matrix), but is an isolated tissue, devoid of blood and lymph vessels,
and nerves.
Cells in articular cartilage - chondrocytes take less
than 10% of the tissue's volume with different zonal arrangement according
to layer of depth. Chondrocytes produce, secrete and maintain the organic
components of extracellular compartment, the organic matrix.
The organic matrix is composed of a dense network (mesh)
of type 11 collagen f brils (10 to 30% by wet weight) enmeshed with proteoglycans
molecules (3 to 10% by wet weight) remaining 60 to 87% is water, inorganic
salts and small amounts of other matrix proteins and lipids.
Coilagen has a high level of structural organization
in articular cartilage, inhomogeneously, but having a layered organization.
Tropocollagen molecules polymerise into collagen fbrils having a diameter
from 25 to 200 nm according to different investigation. The collagen fibre
has tensile stiffness and strength as a basic and most important mechanical
property, but is weak and buckles with compression. Articular cartilage
is anisotropic, its mechanics and material properties differ with the direction
of mechanical loading in relation to the collagen fibers arrangements within
the planes parallel to the joint surface; the number and density variations
in collagen fibers cross-links as well with variation in collagen, proteoglycans
and synovial fluid interactions. Proteoglycans (PG) in cartilage are large
protein-polysaccharide molecules that exist either as monomers or as aggregates.
PG monomers consist of an 200 nm long protein core to which about 150 glycosaminoglycan
chains - GAG (keratin sulfate and chondroitin sulfate - polymer chains
of specific repeating disaccharide units) are covalently attached. The
distribution of GAGs along the protein core is heterogeneous but in arrangement
like a bottle brush structure. PG aggregates are formed from up to 150
PG monomers attached to a central hyaluronate core. Hyaluronic acid core
molecule is to 4 (m long filament of nonsulfated disaccharide chain. Proteoglycan
aggregation promotes immobilization of proteoglycans to the collagen meshwork
and structural rigidity ofthe extracellular matrix.
Water, containing free cations, mostly sodium and calcium,
is the most abundant component of articular cartilage. Concentration of
water decrease with the depth of articular cartilage from 80% on surface
to 65% in the deep zone. It permits difFusion of nutritiens metabolites
between chondrocytes and the surrounding synovial fluid. Only a small percentage
of the water is intracellular, 30% is associated with the collagen fibers
and 70% occupies the intermolecular space and is free to move. This movement
is important for controlling cartilage mechanical behavior and joint lubrication.
Biomehanical behaviour of articular cartilage
Articular cartilage may be considered as a porous permeable
medium (organic solid matrix: collagen fibers and PG mesh) filled with
fluid (synovial fluid).The contact areas in human joints vary according
to joint localization in range of few square centimeters and loading forces
from 0 to several times of body weight. Mechanical behavior of articular
cartilage is viscoelastic and is a combination of the response of viscous
fluid and elastic solid. Two fundamental responses of viscoelastic material
are creep and stress relaxation. With a constant load a viscoelastic solid
as articular cartilage responds with a rapid initial deformation followed
by a slow time-dependent progressively increasing deformation - creep,
until an equilibrium is reached.When articular cartilage is subjected to
the action of constant deformation (strain) high initial stress is followed
by a slow, progressively decreasing of stress (stress relaxation). The
compressive viscoelastic behavior of articular cartilage is due primarily
to the flow of interstitial fluid, and in shear is due to the motion of
long polymer chains - collagen and PGs. During creep the load applied to
the surFace is balanced by the compressive stress developed within the
collagen-PG solid complex and frictional drag generated by the flow of
interstitial fluid during exudation. Creep ceases when the compressive
stress within the solid matrix is sufficient to balance load alone and
no fluid flow and deformation equilibrium is reached. LubricationThere
are several models proposed for lubrication in joints: 1) Fluid film lubrication
model- bearing surfaces are separated by a layer of lubricant. It effect
depends on the reological properties of the fluid. The load on the bearing
surfaces is supported by the pressure in this fluid. 2) Elastohydrodynamic
model and boundary lubrication in which properties ofthe solid surfaces
are combined with those of a thin monolayer of lubricant. Molecules adsorbed
on both bearing articular surfaces. Thickness of these two layers ranged
from 1 to 100 nm (Mow and all 1989, Swann 1985) 3) Weeping lubrication
model in which the articular cartilage is considered as porous, deformable
fluid filled material and, under load, the surfaces were lubricate by a
film offluid expressed from the pores.4) Boosted lubrication model in which
the compression of the articular cartilage traps fluid in irregularities
of articular surfaces. Increasing compression push small molecules through
pores into the cartilage and the synovial fluid on the surfaces becoming
enriched in hyaluronates, more viscous and than more effective lubricant.
Diarthroidal joint surfaces are composed of porous, permeable fluid filed
cartilage that can imbibe and exude a lubricant fluid (Mow et al. 1989.)
Loading and bearing, static and dynamic conditions are extremely variably
suggesting that a mode of lubrication must be a combination of several
mechanisms depending on biomechanical demands.
Friction Friction is defined as the resistance
to motion between two bodies in contact and is defined by coefficient of
friction (. Frictional force (F) is directly proportional to applied load
(W) F = (/W. Coefficient of friction for articular cartilage in synovial
joints was measured between 0.002 and 0.06 (for steel on steel (is 0.6
to 0.8, Ice on ice 0. 1) but is increased with magnitude of the load and
time of application of the load, and is lower when synovial fluid was used
as lubricant than with saline. Wear of articular surfaces According to
Maw and al, wear is the removal of material from solid surfaces by mechanical
action.There are two components of wear: 1) InterFacial wear due to the
interaction of bearing surfaces coming into direct contact with no lubricant
film separating them.There are two ways of this type of wear: a) Adhesive
wear - surface fragments adhere to each other and are torn from the surFace
during sliding. b) Abrasion occurs when a softer material is scraped by
a harder one usually by an opposite bearing surFace or by loose particles
between the bearing. Both adhesive and abrasive wear may take place in
a degenerated joint. Structurally impaired and defected articular cartilage
becomes more permeable and softer.
Lubricant film may leak away through the cartilage
and this loss of lubricant results in direct contact between the asperities
of articular cartilage and exacerbate the removal of cartilage fragments.
2) Fatigue wear results from the accumulation of microscopic damages within
the articular cartilage and subchondral bone under repetitive or cyclically
application of load. Failure may occur over a short period with repeated
high load or over a long period with repetition of low load. (Armstrog
and Mow)
References: Armstrong CG, Mow VC:Variations in
the intrinsic mechanical properties of human articular cartilage with age,
degeneration and water content. J Bone Jt Surg 64A, 88-95, 1982. Armstrong
CG, MowVC: Biomechanics of normal and osteoarthrotic articular cartilage.
In: Clinical trends in orthopaedics. ED:Wilson PD,Jr, Straub LR,Thieme
- Stratton, NewYork 1982. FungYC: Biomechanics, Mechanical properties of
living tissues, SpringerVerlag, NewYork 1981. Junqueira LC, Carneiro J,
Kelley RO: Osnove histologije, Školska knjiga Zagreb, 1995. MowVC, Proctor
ChS, Kelly MA: Biomechanics of articular cartilage. In: Nordin M, Frankel
VH: Basic biomechanics of the musculosceletal system., Lea & Febiger,
Philadelphia, London, 1989. Mow VC, Zhu W, Ratcliffe A: Structure and function
of articular cartilage and meniscus. In: MowVC, HayesWC: Basic Orthopaedic
Biomechanics, Raven Press, NewYork. 199 1. MowVC, Soslowsky LJ: Friction,
lubrication and wear of diathrodial joints. In: MowVC and Hayes WC: Basic
Orthopaedic Biomechanics. Raven Press, NewYork, 1991. NikolićV., Hudec
M: Principi i elementi biomehanike, Školska knjiga, Zagreb, 1988. Swann
DA, et al.: The molecular structure and lubricating activity of lubricin
from bovine and human synovial fluids. Biochem J 225: 195 201, 1985.Williams
PL,Warwick R, Dyson M, Bannister LH: Gray's Anatomy. 37th Ed. Churchill
Livingstone, Edinburgh, London, Melbourne, New York 1989.
The
nosological role of primary necrotizing processes (PNP) in rheumatoid arthritis
Schneider S, Fassbender
HG
Zentrum für Rheuma-Pathologie
(WHO Centre), Mainz, FRG
The profile of RA is characterized by three different components: I.
by immunologic inflammation
2. by an oncologic destruction process
3. by primary necrotizing processes (PNP)
The nosologic significance of PNP hitherto was estimated too low. PNP
only occur in patients suffering from seropositive RA. The following structures
can be affected: Tendon, joint capsule, myocard, artery wall, eye and lung.
The tissue destruction is caused by proteases, released by previously proliferated
fibroblasts. While inflammation and destruction are fimited to joints and
tendon sheaths, PNP are able to manifest at many sites of the organisms,
sometimes fatally, and, under certain circumstances, they can dramatically
influence the nosological picture of RA.
Signal
enhancement method for the digoxigenin-labeled in situ hybridization
Harjaček M, Diaz-Cano
S
Department of Pediatrics,
University Hospital Rebro, Zagreb, Croatia Institute for Pathology, GSF
Forschugscentrum, Neuherberg, Germany
In
situ hybridization (ISH) in tissue sections has a wide range of applications
in biomedical research. However, ISH still remains a time consuming procedure,
and in cases where expression of target mRNA is presumably low, positive
results using a standard NBT/BICP chromogen are often difficult to interpret.
We report a novel, simple, rapid and particularly sensitive method for
detection of the low copy target mRNA. Our new signal detection method
is based on the utilization of biotinylated, monoclonal anti-digoxin-Ab
which cross-reacts with digoxigenin and has not been previously used for
ISH hybridization. We chose to study cytokine expression in various tissues,
in particular IL-4 which is known to have smail transcripts (-650 bps)
and is found only in a small subpopulation of activated T ceIIs. The low
number of expected positive cells coupled with small transcripts should
ensure the development of highly sensitive methodology. As a positive control
for the high copy target mRNA, a riboprobe for oncogene c-myc was also
employed. Our results show that this method is very sensitive, reliable
and quick, and offers some unique features that were not available with
standard color detection methods used previously for the digoxigenin-labeled
ISH.
D.
Ethical forum
- Forensic medicine: Profession? Science? Politics?
Etički forum
- Sudska medicina: Struka? Znanost? PoIitika?
Animal euthanasia
Džaja P, Perić J
Zavod za sudsko i
upravno veterinarstvo, Veterinarski fakultet u Zagebu
Euthanasia in veterine represents human and painless killing of animals.
It should be performed particulary due to economic and medical reasons.
We stress the importance of euthanasia that sholud be performed according
to the Iaw. Therefore in this paper we stressed legislature that determine
the perFormance of euthanasia. In the majority of cases the owners should
decide about euthanasia of their animals, except in those cases that are
regulated by law.
Epidemiology
of suicide in Osijek county
Ugljarević
M, Marcikić M, Badžek S
Department
of Forensic Medicine and Pathology University Hospital Osijek, Osijek,
Croatia
We choose to study suicide in Osijek Country
focusing on trends and levels based on demographic characteristics. Compilation
of most descriptive statistics was performed by a standard statistical
analysis computer package. There were 280 reported suicide cases, among
Osijek Country residents, from 1986. to 1995. The average annual rate was
15,9/100 000 residents from 1986. to 1990., and 18,9/100 000 residents
in the second time period from 1991. to 1995. Male suicides 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
spring and fall, and low figure for January. The suicide rate for male
was the highest for the age group of 25 to 29 year olds and 60-64 year
olds. Among women suicide rate was the highest for the age group of 60
to 64 year olds. Married and retired committed the largest proportion of
suicides. Firearms were preferred by men and poisons by women. There was
a significant rise in the suicide rate by firearms after 1990. Depression
associated with physical incapacitation and intolerable loneliness were
common reasons for suicide among elderly men.
DETERMINATION
OF THE WAR VICTIM'S AGE BY TEETH
Brkić H. Kaić Z, Keros
J, Njemirovskij V
Department of Dental
Anthropology, School of Dental Medicine Zagreb, Croatia
In this paper authors present the the dental age determination methods
of the bodies exhumed from the mass graves in Croatia. The analysis of
teeth is found to be one of the main methods in the identification process
in the following ways:
1. Dental identification achieved by comparison with antemortem data;
2. Identification achieved through an interdisciplinary approach;
3. Identification achieved by the analysis of the DNA isolated from
the dental tissues cells.
When the interdisciplinary approach is used, involving forensic scientists
from different disciplines, the odontologist determinates the age and sex
of the body based on the analysis of teeth and corresponding bones structures.
Dental age has been determined through clinical and x-ray analysis based
on: a) chronology of the teeth eruption; b) quantity of mineralization
of hard dental tissues; c) changes on dental tissues acquired during Iifetime.The
results show that the determination of dental age is a very reliable method
that contributes to faster and precise identifications of the numerous
victims of war.
Literature: Brkić H et al., Dental Identification of War Victims
from Petrinja in Croatia. International Journal of Legal Medicine
1997: 110:47.
Galić J et al., Dentition Based Postmortem Age Identification. Acta
Stomatologica Croatica 1996:29: 197.
Brkić H et al., Dental Identification. Hrvatski Stomatološki
Vijesnik 1997:5:4.
Gustafson G Age Detrminations on Teeth. Journal of American Dental
association 1950:4 1:45.
E.
Computer
systems and computerisation in pathology
Informatički sustavi i kompjutorizacija u patologiji
Thyroid
cancer in clinical thyroid diseases register of University Hospital "Sestre
milosrdnice"
Šimunović D, Belicza
M, Lechpammer M, Gladić V, Kusić Z
"Ljudevit Jurak" Clinical
Department of Pathology, "Sestre milosrdnice" University Hospital, Zagreb,
Croatia
The aim of our research is to make an analysis of the hospital population
through the period of eighteen years (1980- 1997) and to calculate representation
of the thyroid cancer in that sample. We were interested in histological
representation, sex distribution with female to male ratio (F/ M), average
age at diagnosis, presence of metastasis at diagnosis and the size of the
tumor. Following the representation of the follicular carcinoma through
the years we wanted to discover possible abnormalities of the daily iodine
uptake at the examined population. Šeparović et al. claim that in areas
without endemic goiter representation of the follicular carcinoma among
malignant tumors is 5 to 15% while in the areas without an endemic goiter
representation is up to 40%. At the same time we have started creating
the computer register supplemented with a clinical data for further following
of the thyroid cancer. Our intention was to add part of the database in
"Thanatos" computer system that relates to malignant diseases in the Department
of Clinical Pathology "Ljudevit Jurak" to the hospital's Central Register
for Cancer. Biopsy materials were acquired after partial or total thyreoidectomias
of the patients that were worked on in "Sestre milosrdnice" University
hospital. Materials were embedded in paraffin wax, cut in slices 5 (m thick,
colored with hemalaun-eosin and analyzed with microscope. Pathology specialists
analyzed all materials. During the eighteen year period (1980- 1997) at
the Department of clinical pathology "Ljudevit Jurak", University hospital
"Sestre milosrdnice" in Zagreb, there was a total of 196 878 biopsy findings.
We have separated pathohistological diagnosis of the thyroid cancer. With
a total of 379 thyroid cancer biopsy findings. We have found 219 (57,8%
of all cancer) papillary carcinoma, 77 (20,3%) follicular carcinoma, 29
(7,7%) medullar carcinoma, 30 (7,9%) anaplastic carcinoma and on category
other 24 (6,3%) of all thyroid cancer. Findings by other categories of
our interest were: papillary carcinoma had female to male ratio 3,38:1,
metastasis in 26% an average size 1,78 cm (0,73-3,83). Female to male ratio
in follicular carcinoma was 2,21:1, metastasis in 9,1% and average size
2,86 cm (1,5-5). Medullar carcinomas average size was 2,29 cm (0,5-4),
metastasis in 5 1,7 % and F/M ratio 1,42: 1. F/M ratio for anaplastic carcinoma
was 1,73:1, average size 2,75 cm (1,5-5) and presence of metastasis in
33,3%. Average age for papillary carcinoma was 45,4 years, for follicular
cancer 51,1yr., medullar carcinoma 50,7 yr, and anaplastic carcinoma 55,6
yr. Among all thyroid cancers, there has been one difference observed.
Average age of the patients with papillary carcinoma (45,4 yr.) is greaterthan
quoted in Iiterature. The representation ofthe follicular carcinoma among
malignant tumors according to values quoted in literature is 5- 15 % in
areas without an endemic goiter. The representation in our sample is 20,3%
and it is in correlation with the presence ofthe endemic goiter in Croatia.0ur
result suggests further research. Other results are in correlation with
the literature.
Literature: 1 Belgian registry ofthyroid cancer Preliminary epidemiological
characteristics revealed by retrospective study (1988- 1993). Meurisse
M et al., Ann Endocrinol (Paris) 1997. Delisle MJ et al., Ann Endocrinol
(Paris) 1997; 1: 4 13. Park SH et al., Jpn J Clin Oncol 1988; 4: 297
Non-malignant
lesions in clinical thyroid diseases register of University Hospital "Sestre
milosrdnice"
Cvjetko l, Belicza M,
Lechpammer M, Gladić V, Kusić Z
"Ljudevit Jurak" Clinical
Department of Pathology "Sestre milosrdnice" University Hospital, Zagreb,
Croatia
Recent nationwide study showed that 8- 15% of Croatian population has
endemic goiter. The aim of our retrospective study was to make an analysis
of the hospital population during the eighteen year period from 1980 to
1997. We have calculated incidence, sex distribution, and time of the diagnosis
for each of the non-malignant thyroid Iesion. The analysis was done using
the "Thanatos" computer system developed at the "Sestre milosrdnice" University
Hospital. "Thanatos" is a computer data base of the biopsy findings together
with the clinical data. There were total 3552 non-malignant thyroid lesions
during that period.The most common was goiter: 2098 cases or 59,1%.
Among those there were 256 men, and 1842 women with female to male ratio
(F:M) 7,2:1. Average age (AA) of the men was 49,7 yr., and AA of the women
was 46,8 yr.There were 979 adenomas (27,6%): 138 among men (AA=46,7yr),
and 841 among women (AA=44,5yr.) with F:M=6, 1: 1. There were 247 cases
(7%) of the Basedow disease: 23 males (AA=46,4yr.), and 224 females (AA=37yr)
with F:M=9,7:1. Hashimoto disease was found in 156 cases (4,4%): 8 men
(AA=5 1,6yr.), and 148 women (AA=49,8yr.) with F:M= 18,5: 1. Hyperplasia
was the diagnosis in 64 cases (1,8%): 6 men (AA=35,2yr.), and 58 women
(AA=35,3yr.) with F:M=9,6: 1. We have also found five women with the fibroses
thyreoiditis (AA=52yr). More then half or 59, 1% of the non-malignant slesions
goes to goiter. All non-malignant thyroid lesions are more common in women,
especially Hashimoto disease (F:M= 18,51). Most of the non-malignant thyroid
lesions occur during the fourth and fifth decade of life. Hyperplasia is
being diagnosed at the average age of 35,2 yr. in men and 35,3 in women.
Basedow disease in women is being diagnosed at the average age of 37yr
LITERATURE: 1. Kusić Z Endocrinol Invest 1996; 19:204-9. 2.
Žurić M et al. Krušlin B, Robić M. Book of abstracts. The eight "Ljudevit
Jurak" international symposium on comparative pathology. Zagreb, 1997;
33. 3. Park SH et al. Jpn J Clin Oncol 1988; 18:297-302. 4. O'Hanlon DM
et al. Ir Med J 1997; 90: 70.
Causes
of death in autopsies during World War 1 and 80 years later
Kučinar I, Brezovečki
D, Matković M, Lechpammer M
Department of Clinical
Pathology Ljudevit Jurak Clinical Hospital "Sestre milosrdnice"
The main purpose of this study was to investigate major differences
and changes in the causes of death of patients submitted after their death
to our pathologic department but with the time distance of 80 years.
For that purpose we analysed causes of death recorded from September 1916.
to October 1918. in the autopsy reports of the Pathology Unit of Zagreb's
Public Health Institutions (Prosektura javnih zdravstvenih zavoda grada
Zagreba), today's "Ljudevit Jurak" Clinical Department of Pathology. Obtained
data were compared with the causes of death recorded in the archives of
the same institution from January 1995. to December 1997. Memorising and
processing of pathological diagnoses was provided by the "Thanatos" computer
system for clinical pathology.
Comparison of pathologic data originating from 3 war years during World
War I, and 3 years after Croatian war for independence imposes by itself
some significant differences. First, all patients from the 1st investigated
period were male - soldiers died at one of 7 military hospitals existing
in Zagreb at that time. Due to that, 65.5% of them where younger than 49
years, while 29.8% were younger than 29 years of age. Only 2.8% were older
than 50 years at the time of death. In contrast, patients from the 2nd
investigated period were citizens of Zagreb of both sexes died at one of
12 clinical Departments of the Sestre milosrdnice University Hospital.
Their average age at the time of death was 67 years.
However, comparison of these two undoubtedly very different time periods
offers some very intriguing differences in causes of deaths. Among 1118
deaths in the 1st period it was established that infectious diseases were
the main cause of death (60.5%), followed by injuries (12.8%) and respiratory
diseases (10,1%). In the 2nd observed period total of 3538 deaths was recorded.
The main causes of death were diseases of cardiovascular system (23,45%),
followed by respiratory causes (16,6%). It is interesting that remaining
59,95% of deaths is distributed in smaller ratios on wide variety of causes
of death ranging from injuries and malignancies to paediatric deaths.
F.
Stomathological
pathology
Stomatološka
patologija
The effects
of C02 and Nd-yag lasers on the pulp and periodontal tissue on the dog
Jukić S, Anić l, Talan-Hranilović
J
Department of Dental
Pathology, School of Dentistry Zagreb, Croatia
"Ljudevit Jurak" Clinical
Department of Pathology, "Sestre milosrdnice" University Hospita~ Zagreb,
Croatia
C02 and Nd:YAG lasers are most commonly used in medicine and dentistry.
The aim of this study was to evaluate the effect of these two lasers on
dental pulp and peridontal tissue. Mixed breed dogs weight 25 kg were used
for experiments. The animals were anesthetized with pentobarbitural. Pulp
chambers of third premolars and molars were opened and pulp tissues were
removed from it with sharp spoon. The remainder pulp tissue at the root
canal opening of right upper jaws was lased with C02 laser (2 W / 5 tps
/ 10 ms). The teeth in the left sides of the jaws were lased with Nd:YAG
laser (2W / 20 pps). First and second premolars served as positive and
negative controls.The teeth were temporary filled with policarboxylat cement
and procedure was repeated 15 days later on teeth of lower jaws. Peridontal
tissue was exposed by intentional penetration od the pulp chamber floor
with steel bur. After hemostasis and drying the furcation region of upper
right molars was irradiated with C02 laser (2 W / 5 tps / 10 ms) for 3
s. The left side of the jaws was lased with Nd:YAG laser (2 W / 20 pps)
for the same period of time. The same treatment was repeated on teeth of
lower jawes for 15 days. The animals were sacrified by an overdose of pentobarbital
30 days after treating lower jaws and tissue was processed for histopathological
analysis. The most frequent finding in the pulp tissue in case of using
both lasers and regardless oftime oftreatment,was necrosis.A strong chronical
inflammatory reaction without dentin bridge formation over canal of rices
was found.Appearance of more carbonized tissue and changes in deeper portions
of pulp tissue was found in case of Nd:YAG laser irradiation. Fluency that
is used in laser soft tissue surgery causes superficial necrosis, inflammatory
cells' infiltration and in deeper portions osseous transformation. Inflammatory
resorption of cement and cellular infltration of peridontal ligament is
often found beneath superficial wound layer. Nd:YAG laser in case of peridontal
tissue, also penetrates more and causes sever changes of tissue. 45 days
after treatment with C02 laser, first signs of osseous healing response
appeared. In contrast, peridontal tissue lased with Nd:YAG laser did not
show any sign of healing.
Lack
of p53 and nm23-H I genes deletion in oral lichen planus and leukoplakia
Mravak-Stipetić M, Gall-Trošelj
K, Pirkić A, Pavelić K, Pavelić J
School of Dentistry
University of Zagreb, Zagreb, Croatia
Department of Molecular
Medicine Institute "Ruder Bošković", Zagreb, Croatia
"Ljudevit Jurak" Clinical
Department of Pathology "Sestre milosrdnice" University Hospital, Zagreb,
Croatia
In our attempt to identify genetic changes involved in epithelial cell
proliferation, in certain cases as an earliest step in malignant epithelial
neoplasm development, we examined p53(PCR/RFLP) and nm23-H 1(PCR/LOH) allelic
deletions in oral benign , potentially malignant epithelial lesions. The
study group comprised of 25 benign epithelial lesions, clinically diagnosed
as lichen planus (N= 17) and leukoplakia (N=8). Two reccurent aphtous ulcers
as well as one specimen diagnosed a benign migratory glossitis were also
included in the study. Among 21 samples analyzed for exon 4 (p53 gene)
LOH, only 6 (28,5%) were informative with no presence of loss of either
allele. Of 23 samples tested for LOH at intron 6 of p53 gene, 8 (42, 1%)
were informative, again with no presence of LOH. The frequency of alleles
was 73,4 and 26,6% for alleles with and without a specific restriction
site. For nm 23 H- 1gene the analysis was perFormed on total of 24 cases.
Of them 16 (16,6%) were informative, however, non sample exhibited LOH
at his locus. In conclusion, whereas the presence of gross gene alterations
(LOH) would have been definitive evidence for the p53 and/or nm 23 involvement
in hyperproliferation process, the absence of LOH does not exclude the
presence of either smaller mutations, or disregulation of normal gene,
or disfunction at the level of wild type protein. Alternatively, p 53 and
nm 23-H I have no relation to oral lesion formation, and can not be considered
as an early step in benign maybe neoplastic, tissue transformation.
Defence
reaction in a dental pulp after partio-pulpectomy in dog
Pavlica Z, Juntes P,
Pogačnik M
University of Ljubljana,
Veterinary Faculty, Ljubljana, Slovenia
Aim The aim of our work was to study the reactions of a dental
pulp in dogs after radical procedures on teeth. A partial vital pulpectomy
was used as a model.
Material and methods A class V cavity was prepared on
upper third right and left incisors and the dental pulp was exposed.The
same sterile bur was used to create a 2 to 3 mm partial pulpectomy.The
partial pulpectomy wound was treated with calcium hydroxide and covered
with cement and left for standard restoration with composite (1,2,3,4,5).
Partial pulpectomy was performed on 33 teeth. On the 60th day the test
teeth were extracted under anesthesia and prepared for histology. Paraffin
blocks of the decalcifed teeth were oriented to permit buco-lingual sectioning
of the teeth. Eight Nm thick paraffin sections were stained with hematoxylin-eosin
and examined forthe type and degree of pulp inflammation (hyperaemia, pulpitis,
necrosis or gangrene). Degenerative changes of the dental pulp (vacuolation,
calcification, amyloid or hyaline changes) were also determined.
Results and conclusions Inflammation was seen in
one incisor only. A hard continuous barrier with well formed tertiary dentine
and dentinal tubules was seen in all teeth and in all samples it was formed
laterally in a continuous barrier initially. The tertiary dentine differed
from the normal dentine in having fewer tubules, which were strongly curved.
Partial pulpectomy of the dental pulp produced better quality, uninterupted
dentinal bridges in several dogs, when compared to the less radical methods
like direct pulp capping, which we have studied earlier. On the basis of
this results, the radical method of partial pulpectomy was recommended
as the most suitable method for retaining teeth vitality in dogs.
References 1) Ten Cate AR. J Dent Res 1985; 64:549-5 1;
2) Ivanović V et al. Oral Surg 1989; 67:684-8;
3) Schroder U. J Dent Res 1985; 64: 5 1-8;
4) Matsuzaki K et al. Bull Tokyo Dent Coll 1990; 3 1:9- 15;
5) Cullum DR et al. Oral Surg Pathol 1985; 60:218-23.
The
activity of NADPH - d in chronfcalfy fnflammed dental pulp
Jukić S, Sisig S, Anić
I, Mihelić D, Talan-Hranilović J
Department of Dental
Pathology School of Dentistry, Zagreb, Croatia
"Ljudevit Jurak" Clinical
Deparpment of Pathology "Sestre milosrdnice" University Hospital, Zagreb,
Croatia
NADPH - diaphorase serves as an electron donor for nitric oxide (NO)
synthase that converts L- arginin in nitric oxide and citrulin. Recently,
it has been found that nitric oxide may act as a neurotransmitter. This
study was undertaken to determine the activity of NADPH - diaphorase in
human dental pulp with chronical pulpitis and in health controLThirty human
dental pulps were collected out of the routine dental procedure from carious
profunda, nonperiodontaly involved teeth, treatment planed for endodontic
therapy. Health controls were obtained from premolars or third molars extracted
due to orthodontic therapy. Enzyme-activity of human NADPH-diaphorase was
determined by the Pearse et al. (1968) method. High NADPH-d activities
were obtained in pulps with chronical pulpitis.
Transcutaneous
stfmulatfon analgesia fn stomatofogfcal procedures
Kaić Z, Keros-Naglić
J, Božićević D
Department of Dental
Antropology, School of Dental Medicine, Zagreb, Croatia
Pain Control Centre,
Neurosurgical Clinic, School of Medicine, University ofZagreb, Zagreb,
Croatia
The aim of this study was to evafuate the efficiency of transcutaneous
electrostimulation analgesia (TEA) in different stomatological procedures.The
study comprised 180 patients. In 90 of them the stomatological procedure
was carried out under TEA, while the other 90 patients were controls. We
used three points corresponding to the traditional Chinese procedure (Lf-4,
LI-20, ST 6) and WQ I 0-A pulse generator with frequency of 30 and 50 Hz.
Data processing yielded the following results: the stomatological procedure
is less painful when TEA is used.There is no statisticaLly significant
difference of the dental procedure regardless to the use of TEA with respect
to age and sex. The statistically significant difference has been found
between gingival painfulness at the beginning of procedure, than 15 and
25 minutes after in patients under TEA (x2=93,48, df=4, p<0,05). It
has also been found that 58, or 64% of the patients who sustained TEA in
a satisfactory manner, would agree with procedure again. ThereforeTEA may
be considered as an adequate model in study of analgetic value of TEA in
condition of acute pain. Although TEA has no absolute advantage over traditional
pain control procedures, it surely has its indication in patients suffering
from drug allergies, chronic or malignant diseases,AIDS and other progressive
destructive conditions, who cannot be treated with traditional pharmacological
analgesics.
Literature: 1) Turk DC et al. Handbook of pain assessment. NewYork,
Guilford Press, 1992: 15.
2) Lowe NK et al. Confirming the theoretical structure of the McGill
Pain Questionnaire in acute clinical pain. Pain, 199 1;46.
3) Frucht s et al. Muskelentspannung durch transkutane Elektrostimulation
(TENS) bei Bruxismus. Munchen Fortschr Kieferortop. 1995;56.
4) Mumford JM et aLToothache and related pain. Edinburgh, Churchill-Livingstone,
1976; 118.
5) Tabu HA et al. Studies of acupunccture for operative dentistry.
JAD, 1997;95:555.
6) Martin RW et al. Dental dolorometry for human pain research: methods
and apparatus. Pain, 1979; 6:349.
Evaluation
of four methods for estimating taurodontism
Galić J, Kaić Z, Štefanac-Papić
J
Department of Dental
Anthropology, School of Dental Medicine, University of Zagreb, Zagreb,
Croatia
Taurodontism (T) is defined as an apical extension
ofthe pulp chamberwhich results in shortened root. The aims of this study
were to evaluate four methods of "T" estimation and determination of its
concordance.The number of 221 lower molars were traced from the orthopantomograms
of 87 males 24-3 1yr of age. Seven variables were measured on the tracings:Variable
1) lowest point of the roof - highest point of the floor of the pulp chamber,
2) lowest point of the roof of the pulp chamber - apex of the longest root,
3) baseline connecting the two cementoenamel junctions (neck) . highest
point of the floor of the pulp chamber, 4) neck - deepest pit of the occlusal
surFace, 5) neck - root bifurcation, 6) root bifurcation - deepest pit
of the occlusal surface, 7) root bifurcation - apex of the longest root.
Each tooth was assessed twice by the same author (J.G.) within 2-4 days.
T was estimated according to the methods proposed by Shifman (1),Tulensalo
(2),Jorgenson (3) and Seow (4).The value ofVariable 3 proposed by Shifman
was adjusted for magnification on the orthopantomograms while the index
proposed by Seow was adjusted for the second and third molars. Intraexaminer
reliability in reproducing the same classification was, on average, 96,84%,
the highest for Shifman's method (99,10%) and the (owest for the Seo~n%s
method (94,57%).Table 1 shows frequencies of taurodontism found and methods
used.Table 1The concordance in diagnosingT was the highest for Shifman's
and Jorgenson's method (94, 1%) and the lowest fofTulensalo's and Seow's
method (83,8%). According to Shifman's criterion only two molars were classified
as taurodontic and those were the only teeth thet were classified the same
by all four methods.Thus, the method for estimatingT described by Shifman
proved to be the most reliable for use in later epidemiological investigations.
References: 1) Shifman A. et al. Community
Dent Oral Epidemiol 1978; 26:200.
2)TulensaloT et al. Community Dent Oral Epidemiol
1989; 17:258.
3) Jorgenson R.J. et al. J Cran Genet Dev Biol
1982; 2: 125.
4) Seow W K. et al. Pediatric Dentistry 1989;
11(3):2 14