Correlation between phenotypic characteristics of mononuclear cells isolated from human periapical lesions and their in vitro production of Th1 and Th2 cytokines.

UNLABELLED: The development and progression of periapical dental lesions, mediated by the specific immune response, are poorly understood. In these processes, an interplay of different proinflammatory and immunoregulatory cytokines is of crucial importance. OBJECTIVES: To examine the activation of T helper 1 (Th1) and Th2 immune responses in 25 human periapical lesions based on the ex vivo production of interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) by mononuclear cells (PL-MNC). METHODS: The levels of IFN-gamma and IL-4 in culture supernatants of PL-MNC, determined by ELISA, were correlated with concentrations of these cytokines in cultures of control MNC from peripheral blood (PB-MNC), cellular composition of inflammatory cells and phenotypic characteristics of PL-MNC. RESULTS: We detected high levels of IFN-gamma in all samples, after cell stimulation with phorbol myristate acetate and Ca(2+) ionophore, that were not statistically different from the levels of IFN-gamma in PB-MNC cultures. IL-4 was detected in 76% samples, but its concentrations were much lower than in PB-MNC samples. The levels of IFN-gamma were higher in cultures of PL-MNC isolated from periapical lesions with predominance of T cells (T-type lesions) and correlated positively with the proportion of antigen-presenting cells (macrophages and dendritic cells), CD4(+) T cells and IgG2(+) B cells/plasma cells. The levels of IL-4 correlated negatively with the proportion of macrophages, but positively with the number of mast cells and IgG4(+) cells. IL-18Ralpha, a stable marker of Th1 cells, was detected on a relatively small proportion of CD3(+) T cells and its expression correlated with the levels of IFN-gamma. However, the expression of ST2L, a stable Th2 cell marker, was not detected. The levels of Th1 and Th2 cytokines did not correlate with clinical characteristics of the lesions, defined by the presence of symptoms. CONCLUSION: Cumulatively, our results suggest the predominance of Th1 immune response in periapical lesions.

[Significance of pathohistological findings and the expression of Bcl-2 in diagnosis and treatment of oral planocellular carcinoma].

BACKGROUND/AIM: Numerous studies were aimed to detect and characterize various tumor markers in patients with oral planocellular carcinoma in order to reduce moratlity and mobidity rates of these patients, as well as to establish the correlation between the expression of specific tumor marker and prognostic outcome. The aim of this study was to determine patohistological characteristics of tumor and peritumor tissue in patients with oral planocellular carcinoma, with special regard to the expression of Bcl-2, as well as to point out the significance of clinicomorphological correlations for clinical use. METHODS: Sixty-two patients with oral planocellular carcinoma, stage II and III, were examined. The patients were surgically treated for this condition at the Clinic for Maxillofacial Surgery, Military Medical Academy, Belgrade. Surgical specimens were obtained from both tumor and peritumoral tissues. Patohistologic degree of tumor differentiation and the immunohistochemical expression of Bcl-2 were determinated for each specimens. RESULTS: Twenty-four (39%) patients had tumor dimension T1, while six (9%) and thirty-two (52%) patients had tumor dimension T2 and T3, respectively. Patohistologic analysis of peritumor connective, fat, muscle and bone tissue samples confirmed the presence of tumor infiltration. The expression of Bcl-2 in peritumor tissue samples correlated significantly with tumor's histologic grade (rho = 0.468; p < 0.001), nuclear grade (rho = 0.430; p < 0.001) and nucleocytoplasmic ratio (rho = 0.410; p = 0.001). CONCLUSION: This results suggest that the expression of Bcl-2 in combination with patohistologic findings could have a prognostic value in patients with oral planocellular carcinoma.

[Ambient temperature impact on hepatocellular liver damage in rats following intake of 3,4-methylenedioxymethamphetamine].

BACKGROUND/AIM: 3,4-methylendioxymethamphetamine (MDMA, Ecstasy) is a psycho-stimulating agent. It is usually taken orally in the form of tablets. It is absorbed throught the gastrointestinal mucous membrane. Hyperthermia is the most prominent clinical sign of MDMA intake. The most prominent forensic finding of lethal MDMA poisoning is myocardial infarction and cerebrovascular bleeding. However, liver and kidney damage: have also been described. The aim of this research was to determine if ambient temperatures affect liver damage in the experimental rats. METHODS: The experiment was conducted for 8 h and 24 h, at temperatures of 12 degrees C, 22 degrees C and 32 degrees C. Both biochemical parameters (ALT, AST, AP, gamma GT and LDH) and pathohistological changes of the liver were monitored. RESULTS: Our reserch demonstrated that the most serious lever damage occurred at 32 degrees C. Liver damage was manifested as portal inflammation, periportal necrosis, lobular necrosis, stasis, intralobular hemorrhage and incerease of liver enzymes serum activity. CONCLUSION: Liver damage after MDMA intake rises with the increase of ambient temperature, and it is most pronounced at the temperature of 32 degrees C.

[Investigation of Balkan endemic nephropathy in Serbia: how to proceed?].

Balkan endemic nephropathy (BEN) presents an unsolved puzzle despite fifty years of its investigation. Academy of Medical Sciences of the Serbian Medical Society organized a round table discussion on current unsolved problems related to BEN. The present paper summarizes presentations, discussion and conclusions of this meeting. During the last fifty years, the course of BEN prolonged and it shifted towards the older age in all endemic foci. Data on the incidence of BEN have been controversial and frequently based on the data on the number of BEN patients starting haemodialysis treatment. In Serbia, BEN patients present 6.5% of haemodialysis population and this percentage differs among different centres ranging from 5% (Leskovac) to 46% (Lazarevac). Maintenance of high prevalence of BEN patients on regular haemodialysis indicates that BEN is not an expiring disease. In addition, recent data have shown more frequent microalbuminuria and low-molecular weight proteinuria in children from endemic than from nonendemic families. Aetiology of BEN is still unknown despite numerous investigations of environmental and genetic factors. Today, there is a very current hypothesis on the aetiological role of aristolochic acid but the role of viruses, geochemical factors and genetic factors must not be neglected. Morphological features of BEN are nonspecific and characterized by acellular interstitial fibrosis, tubular atrophy and changes on pre- and postglomerular vessels. New immunohistochemical and molecular biology methods offer a new approach to BEN investigation. Association of BEN with high incidence of upper-urothelial tumours is well-known. Recent studies have shown significant changes of demographic characteristics of patients suffering upper-urothelial tumours, their prevalence in different endemic foci and characteristics of tumours. Further studies of BEN should be directed to determination of incidence and prevalence of disease in different endemic foci, investigations of different insufficiently examined aetiological factors as well as pathomorphological features of the disease by the use of modern methods.

Asymptomatic urinary abnormalities: histopathological analysis.

The aim of the study was to assess the characteristics of histopathological changes in 120 young males, both recruits and soldiers, who had undergone successful renal biopsy due to asymptomatic urinary abnormalities. The patients were subdivided into a group with isolated microhematuria (IMH-62 patients) and a group with asymptomatic microhematuria and proteinuria (MHP-58 patients). Light, immunofluorescence, and electron microscopy revealed that MHP was associated with more severe morphological changes, than IMH. The latter group included 6 subjects with normal biopsies and 13 subjects with minor abnormalities found only in two patients with MHP. The frequencies of particular nephropathies in the groups with IMH and MHP were as follows: 35% and 55% for IgA nephritis, 24% and 31% for non-IgA mesangioproliferative glomerulonephritis (GN), 2% and 3% for focal proliferative GN, 3% and 3% for diffuse proliferative GN, 5% and 1% for thin basement membrane nephropathy, respectively. Rebiopsy, performed in eight patients due to worsening of proteinuria during the follow-up period, showed evidence of progression of morphological changes. Patients with IMH had significantly less prominent histopathological changes than patients with MHP. Therefore, renal biopsy cannot be recommended for patients with IMH unless specific indications are present.

Effect of carvedilol on pulse pressure and left ventricular hypertrophy in spontaneously hypertensive rats with adriamycin nephropathy.

Recent studies indicated pulse pressure as a risk factor for left ventricular hypertrophy, myocardial infarction, congestive heart failure and stroke as well as chronic renal failure progression. The present study examined the effects of carvedilol and its combination with captopril on blood pressure, left ventricular hypertrophy, kidney vascular changes and kidney function in spontaneously hypertensive rats with adriamycin nephropathy. Four groups of 20 SHR each were involved: (1) control group: SHR; (2) ADR group: SHR treated with ADR (2mg/kg i.v. twice in 20 days); (3) ADR-C group: SHR treated with ADR and carvedilol (30 mg/kg/day) and (4) ADR-CC group: SHR treated with ADR and carvedilol (30 mg/kg/day) and captopril (60 mg/kg/day). Systolic-, diastolic- and mean-pressures and pulse pressure were determined at weeks 6 and 12 after the second ADR injection; and body weight, creatinine clearance and proteinuria at weeks -3, 6 and 12. The rats were sacrificed at week 6 or 12, the weights of the left and right ventricles and kidneys measured and the kidney vascular index was calculated as described by Bader and Mayer. Both carvedilol alone and combined with captopril significantly reduced systemic blood pressure but the effect of the latter was more pronounced and registered from week 4 till the end of the study. Carvedilol and its combination with captopril significantly decreased SBP, DBP and MAP. They also decreased PP, prevented the development of LVH, and renal vascular changes and slowed the progression of chronic renal failure and these effects were stronger in the ADR-CC group than in the ADR-C group. The antihypertensive drugs failed to prevent proteinuria in ADR SHR. Significant positive correlations were found between PP (but not SBP, DBP and MAP) and both proteinuria and Ccr in all groups of rats. In conclusion, carvedilol alone, but more strongly in combination with captopril, significantly reduced blood pressure, PP, LVH, renal blood vessel changes and chronic renal failure progression.

[Alport's syndrome and benign familial haematuria: light and electron microscopic studies of the kidney].

INTRODUCTION: Hereditary nephropathy is clinically characterized by the familial occurrence in successive generations of progressive haematuric nephritis and neural hearing loss. Hereditary nephropathy of Alport's syndrome (AS) and benign familial (recurrent) haematuria (BFH) are morphologically characterized by specific and diagnostically important thickening and splitting of lamina densa of the glomerular basement membranes. Those lesions can be recognized only by electron microscopy. Hereditary nephritis is usually present clinically with haematuria, and new mutations without a family history of haematuria. It is therefore important to differentiate hereditary nephritis from BFH and no familial haematuria. Thus, electron microscopy is essential in diagnosis of haematuria. OBJECTIVE: The aim of this study was to describe, by light microscopy, constellation of renal alterations by which hereditary nephropathy can be recognized with high probability as well as to compare the diagnostic validity of the findings observed by light and electron microscopy in AS and BFH. METHOD: We examined 48 renal biopsies of the patients with hereditary nephoropathies by light and electron microscopy. Tissue samples were fixed in buffered paraformaldehyde and embedded in paraffin for long-term preservation. For the electron microscopy analysis, the following fixation in 4% glutaraldehyde tissue was postfixed in 1% osmium tetroxide. Thereafter, the following dehydration procedure tissue slices were embedded in epon. RESULTS: Our results demonstrated that the interstitial foam cells, foetal-like glomeruli, minimal glomerular abnormalities with stain less intense in basement membranes, mild irregular mesangial widening, focal thickening of Bowman's capsule, foci of dilatation tubules, tubular ectasia and atrophy, erythrocyte tubules casts were present in hereditary nephritis. Additionally, light microscopic biopsy findings in patients with BFH were either normal or revealed minor changes (e.g. increased mesangial matrix). All biopsies were reevaluated by electron microscopy and ultrastructural findings confirmed the diagnosis of hereditary nephropathies. CONCLUSION: The findings observed by light microscopy represent an important step that leads to a definitive diagnosis of AS and BFH.The definitive diagnosis, however, depends on electron microscopy.

[Comparison of the suppressing capecitabine and 5-fluorouracil effects on the pronounce of inflammatory cells score in the induction of episcleral fibrosis after trabeculectomy].

BACKGROUND/AIM: Trabeculectomy is a filtering antiglaucoma surgery performed to facilitate the formation of a drainage channel for the aqueous humor from the anterior chamber of the eye to the subconjunctival space making a collector, filtering cushion of the eyeliquid. The cause of unfavourable outcome of trabeculectomy is episcleral fibrosis which leads to the obstruction of the drainage channel Inflammatory reaction is an initial, but, also, the key event in the process of fibrosis and directly proportional to it A successful medicamentous suppression of inflammation could stop fibrosis. Arccordingly, the aim of this study was to comparatively evaluate suppressing effects of capecitabine and 5-fluoroaracil upon inflammation, an initial fibrosis process after trabeculectomy. METHODS: This study was performed on an experimental animal model, namely 2 4 rabbits (48 eyes), which were exposed to artificial cause of the tissue changes analogous to human subclinical inflammation. During the trabeculectomy, a 5% solution of 5-fluoroaracil was applied to the right eye sclera of a rabbit, while 2% solution of kapecitabine was applied to the left eye sclera, thus making possible to evaluate the effects of different drugs to the same animal. Two weeks after the surgery the subconjunctival - episcleral tissue cuts were analyzed. Semiquantitative histologic test was used to compare the cellular elements of inflammatory (fibroblasts, mononuclears, plasmocytes, eosinophiles), as well as fibrosing (colagen, reticulin fibers) process, and, based on the differences in histopathologic score, suppressive efficacy of 5-fluoroaracil and kapecitabine was evaluated and compared between the groups of the treated tissues. The results were shown as mean values +/- standard deviation. RESULTS: A statistically significantly lower score was revealed in the initial inflammatory cells - mononudears and plasmocytes in the group of the tissues treated with kapecitabine, while there was no differences between the groups in the score of fibroblasts and colagen fibers. CONCLUSION: The suppression of the initial episcleral fibrosis process obtained by kapecitabine was more significant as compared to the suppression obtained by 5-fluoroaracil, thus providing more favorable outcome of the trabeculectomy.

[Clinical pathology].

This work describes the basic elements of pathology used in clinical practice. Pathology plays an important role in clinical and scientific work, but only a few areas of pathology will be covered. Although the contribution of oncological and surgical pathology to therapy is the most well known, the cases chosen here will involve infectious pathology, diseases of the kidney and the liver, autoimmune diseases, as well as organ transplantation. Especially important is the description of methods that enable more accurate morphological diagnoses, such as histochemistry, immunohistochemistry, immunofluorescence, and electronic microscopy. Previous experience and joint work with clinical doctors have enabled the definition of significant morphological elements as well as of essential methods of pathohistological diagnosis. Besides, as is often the case, although disease symptoms are difficult to discern and biochemical results do not show significant changes compared to normal values, the results of biopsy come as a surprise to clinical doctors. For example, in virus hepatitis B involving so-called asymptomatic HBsAg carriers, we discovered every morphological form of hepatitis, from minimal lesions to chronic, persistent, and active hepatitis. With hepatitis C, certain morphological lesions point to the etiopathogenesis of this disease and thus help to confirm the diagnosis and to instigate therapy on time. Another significant experience involves kidney biopsies in cases when clinical findings are asymptomatic. Often, in such cases, morphological findings point to glomerulonephritis and glomerulopathy at different stages. Timely and subtle morphological diagnostics offer a more precise explanation for the pathological injury of tissues than other diagnostic methods. In this way, by adopting new methods, the work of pathologists is included more and more in everyday clinical practice. The inclusion of pathologists in a transplantation team makes sure a proper selection of the organ for transplantation is carried out and ensures a reliable evaluation of the condition of the transplanted organ, enabling appropriate therapy. Autoimmune, hereditary diseases are almost impossible to recognise unless a biopsy is performed as in the examples given. In this work, the 30-year-long results of the cooperation between clinical doctors and pathologists are presented and compared with similar results from modern literature, together with numerous examples that represent significant experiences and achievements of our medicine.

[Occurrence of autoimmune chronic hepatitis in siblings].

BACKGROUND: Autoimmune hepatitis is a chronic inflamatory disease of the liver of unknown etiology, characterized by the loss of tolerance against hepatic tissue, leading to the destruction of hepatic parenchyma. It predominantly affects females, and rarely occurs in the same family. CASE REPORT: In this paper we presented brother and sister with autoimmune hepatitis according to the criteria of the International Autoimmune Hepatitis Group. CONCLUSION: Because of a possible genetic predisposition to autoimmune hepatitis, the occurrence of the disease in a family member suggested the need to examine other family members.

[Acute strong headache in emergency neurology (diagnostic and treatment)]. / Akutna, jaka glavobolja u urgentoj neurologij (dijagnostika i lijecenje).

Acute, strong headache represent a frequent symptom with patients who report to the Emergency Neurology. Most of them have functional headaches from the group of primary headaches. However, a smaller number may have secondary headaches, due to structural changes of the cerebral parenchyma. Though the second group is considerably smaller, the secondary headaches, besides fast diagnostics require fast therapy, since they frequently imperil directly the patients life. The paper consider clinical and neurological predictions, indicating possible secondary headaches ("red flags") and diagnostic procedures, necessary to achieve their fast ethiological diagnosis. We consider also possibilities and limitations of certain additional methods (CT, cerebrospinal fluid examination, MR) with the aim to establish the examinations algorithm. Finally we offer procedures related to the therapy of acute primary as well as secondary strong headaches.

[Significance of iron reduction for the therapy of chronic hepatitis C]. / Znacaj uklanjanja gvozda u terapiji hronicnog hepatitisa C.

BACKGROUND: It has been established that many patients with chronic hepatitis C have elevated serum iron, feritin levels and iron deposites in the liver. Therefore, the liver damage due to hepatitis C virus may be aggravated with iron overload. In many studies higher levels of iron in the blood and the liver were connected with the decreased response to interferon-alfa therapy for chronic viral hepatitis C. Recent introduction of pegylated interferons plus ribavirin has improved the therapeutic response, so it is now possible to cure more than 50% of the patients. CASE REPORT: Three patients with chronic hepatitis C and iron overload were presented. Iron reduction therapy using phlebotomy or eritrocytapheresis with plasmapheresis was done at different times in regard to specific antiviral therapy or as a sole therapy. CONCLUSION: It has been shown that iron reduction, sole or combined with antiviral therapy, led to the deacreased aminotransferase serum activity and might have slown down the evolution of chronic hepatitis C viral infection.

[Urgent neurologic states: experience at the Neurology Clinic in Sarajevo]. / Urgentna stanja u neurologiji: iskustva Neuroloske klinike u Sarajevu.

There is a quite good definition of medical care for patients suffering from chronicle neurological diseases. However the neurologist role in taking care of urgent cases is substantially less determined. This paper is analyzing one year efforts of the on duty neurological team in the Out Patient Department and Emergency Division of the Neurology Department in Sarajevo. During this period the on duty neurological team examined the total of 3939 patients, out of which 1022 patients where kept for treatment. The patients where most frequently assigned to the Emergency unit for following reasons: vascular incident of the Central Nervous System(1955 patients or 50%), cerebrovascular accident represented with 1290 or 33%, and TIA of the carotid and vertebrobasilar area 544 or 14% along with hypertensive encephalopathia, 118 or 3%. This is followed by the group of the short-term disturbance of consciousness (472 or 125), out of which the consciousness crises represented 257 or 7%, and epileptic crises 215 or 5%. Following are the lower percentages of the headaches (287 or 7%), radicular painful syndrome of cervical and lumbal area (209 or 5%), vertigo (183 or 5%), neurophatia (167 or 4%), etc. The more extensive number of patients admitted at the Emergency Division where suffering from brain stroke (800 or 78%), TIA was represented by a lower number (172 or 17%). Only 50 patients had other diagnosis. The ischemic stroke represented 674 or 81% with patients suffering from the brain stroke and the hemorrhagic stroke 153 or 19%. Today, the urgent neurological conditions represent a particular area of Neurology, not only neurologists need to know but also other medical doctors, to enable the patients to be forwarded on time to the appropriate care institution.

[Advanced prostatic carcinoma with low levels of serum prostate-specific antigen]. / Uznapredovali karcinomi prostate sa niskim serumskim vrednostima prostata-specificnog antigena.

The serum levels of prostate-specific antigen (PSA) represent a significant diagnostic and monitoring parameter of prostatic carcinoma (PC). The aim of the study was to establish correlation of serum PSA level in addition to grade, histological type, and clinical stage of PC in patients with normal or intermediary PSA serum level. In 37 untreated PC patients with preoperative serum PSA levels ranging between 0.1 and 9.6 ng/ml, paraffin-embedded tissue and serum samples were immunohistologically studied and immunoassay for PSA was done. The most representative was poorly differentiated PC with D stage. In serum samples from PC patients 27 (73.7%) normal (< or = 4.0 ng/ml), and 10 (27.3%) intermediate (4.1-10 ng/ml) PSA levels were found. Immunohistochemistry, in 36 PC (97.3%) had demonstrated the expression of PSA. Our study results had shown low serum PSA levels in some patients with advanced poorly differentiated PC.

[Role of various virus genotypes in progression of chronic hepatitis C]. / Uloga razlicitih genotipova virusa u progresiji hronicnog hepatitisa C.

The relation between HCV genotypes and the progression of chronic hepatitis is still unknown. Some studies implied more pathogenic effect of genotype 1b for the severity of liver inflammation. However, other studies did not show the association between HCV genotype 1b and the severe outcome of HCV infection. The aim of this study was to determine the most frequent genotypes in this environment and their influence on hepatitis C severity. The investigation included 34 patients with histologically confirmed chronic hepatitis C, aged 20-65 (mean 35.0 years). On the basis of pathohistological findings, applying the modern classification, the disease activity was graded as: minimal (A1) moderate (A2) and severe (A3). The extent of fibrosis was marked as: absent (F0), mild (F1), moderate (F2) and severe (F3). Genotyping was performed by nested PCR with type-specific primers and LIPA test and verified by sequencing. The most prevailing genotype in our group of patients was 1b (44.1%), followed by genotype 3a (26.4%), genotype 1a (11.7%) and 2a (2.8%). Five patients had mixed genotypes (four 3a/1b, and one 1a/1b). The severity of liver cell necrosis, measured by alanintransferase (ALT) levels in serum was not related to any of HCV genotypes. There was no statistically significant difference between histological disease activity in relation to HCV genotypes. Stage of the disease was not significantly related to the HCV genotypes. There was a strong association between the degree of fibrosis and the age of patients (p < 0.01). These results could indicate that the determination of HCV genotypes was not useful in the estimation of disease severity and that liver biopsy was the most important for the prognosis of the disease.

[Actinomycosis--a multidisciplinary approach to a clinical problem]. / Aktinomikoza--multidisciplinarni pristup klinickom problemu.

Actinomycosis is caused by microorganisms of the Actinomyces species, found in the oral cavity, gastrointestinal and genital tract. Infection by Actimomyces species mainly causes chronic supurative disease of the oral, cervico-facial, thoracic, and abdominal regions. Clinical manifestations have been described at every level, but the disease is relatively rarely diagnosed. The causative agent is considered to be an opportunistic pathogen associated with infection, trauma, surgical procedures, or associated with a neoplasm or foreign body. Symptoms include intermittent fever, weight loss, abdominal or thoracic pain, pseudotumor and tumor masses, and local symptoms from the infected area. Pseudotumor and tumor masses are diagnosed by ultrasonography and computerized tomography. The diagnosis is confirmed by the presence of a causative agent in biopsy or surgically acquired samples. The treatment includes the administration of high doses of antibiotics (penicillin, amoxicillin, ceftriaxone) for at least 6-12 months, as well as surgical treatment. Penicillin is a drug of choice. This infection is often polymicrobial, so that antibiotic therapy should include agents that act against the associated pathogens. Treatment outcome and prognosis in these three cases were excellent, despite the prolonged course of the extensive disease.

[Value of urinary cytology findings in the diagnosis of acute renal graft rejection]. / Vrednost nalaza citoloskog pregleda urina u dijagnozi akutnog odbacivanja transplantovanog bubrega.

BACKGROUND: Acute rejection of allograft is one of the most serious complications of renal transplantation that requires fast and precise diagnostic approach. In this paper our experience in cytologic urinalysis as a diagnostic method of the acute renal allograft rejection was reviewed. METHODS: The study group included 20 of 56 patients with transplanted kidneys who were assumed for the acute allograft rejection according to allograft dysfunction and/or urine cytology findings. Histological findings confirmed allograft rejection in 4 patients. Urine sediment obtained in cytocentrifuge was air-dried and stained with May-Grunwald-Giemsa. Acute allograft rejection was suspected if in 10 fields under high magnification 15 or more lymphocytes with renal tubular cells were found. RESULTS: Acute transplant rejection occurred in 32.1% patients. In 15 patients clinical findings of the acute renal allograft rejection corresponded with cytological and histological findings (in the cases in which it was performed). Three patients with clinical signs of the acute allograft rejection were without cytological confirmation, and in 2 patients cytological findings pointed to the acute rejection, but allograft dysfunction was of different etiology (acute tubular necrosis, cyclosporine nephrotoxicity). In patients with clinical, cytological and histological findings of the acute allograft rejection urine finding consisted of 58% lymphocytes, 34% neutrophilic leucocytes and 8% monocytes/macrophages on the average. The accuracy of cytologic urinalysis related to clinical and histological finding was 75%. CONCLUSION: Urine cytology as the reliable, noninvasive, fast and simple method is appropriate as the a first diagnostic line of renal allograft dysfunction, as well as for monitoring of the graft function.

[Importance of cytologic examination of urine in the diagnosis of renal transplant function disorders]. / Znacaj citoloskog pregleda urina u dijagnostici poremecaja funkcije transplantovanog bubrega.

BACKGROUND: This paper presents our experience with cytologic examination of urine in diagnosing renal allograft dysfunction. METHODS: The study group included 23 patients with renal allograft dysfunction, selected from 56 patients who underwent renal transplantation. Etiologic diagnosis was made according to the clinical picture, histological findings during allograft biopsy, and cytologic examination of urine. Urine sediment was obtained in cytocentrifuge and was air dried and stained with May Grunwald Giemsa. RESULTS: Out of 23 patients with allograft dysfunction in 18 (78.3%) patient it was caused by acute rejection, and in 5 (8.9%) patients by allograft infarction, cyclosporine nephrotoxicity, acute tubular necrosis and chronic nephropathy. In eighteen patients (78.3%) cytologic examination of urine was pathologic, while in 16 (70%) clinical and histology findings coincided with urine cytology findings. Out of 18 patients with acute allograft rejection in 15 patients cytologic examination of urine coincided with acute rejection. Out of 7 patients with expressed cyclosporine nephrotoxicity, in 5 cytologic examination of urine confirmed the cause of allograft dysfunction, as well as in one of 2 patients with acute tubular necrosis. Cytologic examination of urine indicated parenchymal damage in 2 patients with recurrent disease (membranoproliferative and focal sclerosing glomerulonephritis). In 4 of 5 patients suffering from chronic rejection in a year's monitoring period, urine sediment periodically consisted of lymphocytes, neutrophilic leucocytes, monocyte/macrophages, tubular cells and cylindres, without the predominance of any cell type. In 3 patients allograft dysfunction was caused by infective agents (bacteria, fungus, cytomegalovirus). CONCLUSION: Cytologic examination of urine might be an alternative to histological in diagnosing acute allograft rejection and acute tubular necrosis or nephrototoxicity. Also it might indicate parenchymal disease while the importance of urine cytology in chronic allograft nephropathy needs to be investigated further.

[Slowing the progression of chronic renal insufficiency with captopril in rats with spontaneous arterial hypertension and adriamycin nephropathy]. / Usporavnje progresije hronicne insuficijencije bubrega kaptoprilom kod pasova sa spontanom arterijskom hipertenzijom i adriamicinskom nefropatijom.

INTRODUCTION: The role of hypertension in chronic renal failure (CRF) progression was described in 1914 by Volhard and Fahr [1], in 1940 by Rite and colleagues [2] and subsequently many studies described the effects of various antihypertensive drugs on regulation of blood pressure and CRF progression. The recent experimental and clinical studies especially emphasized the role of angiotensin converting enzyme (ACE) inhibitors in the regulation of hypertension and slowing down of CRF progression, but there are still issues for discussion and disagreement [2-14]. The aim of this study was to analyse the effects of captopril on clinical, biochemical and morphological changes in spontaneously hypertensive rats (SHR) with adriamycin (ADR) nephropathy. SUBJECTS AND METHODS: Experimental animals. Adult (24 weeks) female spontaneously hypertensive rats (SHR), weighting about 200 g, were bred at the Institute of Medical Research, Belgrade. The rats were randomly divided in the following groups: 1. CONTROL GROUP: 12 SHR; 2. Adriamycin group (ADR): 27 SHR treated with adriamycin (2 mg/kg i.v. twice for 20 days); 3. Adriamycin-captopril group (ADR-C): 30 SHR treated with adriamycin and thereafter with captopril (60 mg/kg/day). Animals were followed-up for 18 weeks after second adriamycin injection. Systolic blood pressure was measured at 2 weeks intervals throughout the study. Blood and urine samples were collected in weeks--4, 6, 12, and 18. Morphologic studies. Rats were killed at weeks 6, 12 or 18 after the second adriamycin injection, when the kidneys were removed and fixed in neutral buffered formalin (10%). Paraffin embedded tissue sections 4 microns thick were stained with hematoxylin and eosin, periodic acid-Schiff reagent (PAS), Thrichrom Masson and Silver methanamin (Jones) for light microscopic study. A semiquantitative score was used to evaluate glomerular, vascular and tubulointerstitial changes. A minimum of 60 glomeruli for each kidney were examined, and the severity of the lesions was graduated according to the percentage of glomerular sclerosis from 0 to 10 (0--0%; 2--20%; 5--20-50%; 10--100%) [16]. Vascular changes were graduated from 0 to 3 according to hyalinosis in the walls of the artenoles (1--0%; 2--< 50%; 3--50-100%; 4--100%) [17]. Tubulointerstitial changes were semiquantitatively expressed by calculation of separately the index of interstitial fibrosis (0--0%; 2--< 20%; 5--> 20%; IQ--> 40%) and the index of interstitial infiltration and tubular atrophy (0--0%; 1--< 20%; 2--> 20%; 3--> 40% [18]. Results were presented as mean +/- SD. Differences between groups in functional data as well as morphologic lesions were studied by one-way analysis of variance and the unpaired T-test. RESULTS: Captopril decreased systemic blood pressure in ADR SHR significantly, but failed to prevent proteinuria (Fig. 1). Urea and creatinine in serum progressively increased in all studied groups, but faster in ADR SHR groups than in controls (Table 1). Creatinine clearance decreased faster in ADR group than in ADR-C group, but without statistical significance (Table 1). Among sixty nine analysed rats at the beginning of the study, sixteen died during the study. The other animals (Table 1) were killed at weeks 6, 12 and 18; pathohistological changes of their kidneys with glomerular, vascular and tubulomterstial indexes are presented in Table 2. In control group of rats minimal glomerular and interstitial changes could be seen in week 18, mild tubular changes were present in weeks 12 and 18, and marked changes in blood vessels were manifested in week 12, as well as in week 18 (Fig. 2, Table 2), when their statistical significance was higher than in rats treated with adriamycin. Glomerular, tubular and interstitial changes were mostly pronounced in adriamycin treated rats and became more expressive during the experiment (Table 2, Fig. 3). In ADR-C group of rats captopril slowed down glomerular changes, but significantly in week 18 only (Table 2). The same was with interstitial changes (Table 2, Figs. 3-c, 4-c). Tubular and vascular changes were less in week 6 in ADR-C group than in ADR group, what was leveled off later in the study (weeks 12 and 18) (Table 2). DISCUSSION: Although Richard Bright was probably the first person to notice that severe renal diseases were associated with changes of the cardiovascular system, Volhard and Farhr first described that high blood pressure was the major cause of progressive loss of renal function in chronic renal diseases [1]. Subsequently, many authors in their experimental and clinical studies described the effects of various antihypertensive drugs on regulation of blood pressure and slowing down of CRF progression. Various experimental models were used in their studies [8, 19-21]. With discovery of ACE inhibitors and first studies which pointed that this group of drugs effectively slowed down CRF progression, many authors studied their effects on systemic blood pressure regulation, reduction of glomerular hypertension and slowing down of CRF progression. Anderson, Rennke and Brenner studied the effects of "triple therapy" (reserpine, hydralazine and hydrochlorotiazide) and ACE inhibitor enalapril in rats with subtotal nephrectomy [3]. Enalapril decreased systemic blood pressure, glomerular hypertension, proteinuria and glomerular sclerosis [9, 22-24], while "triple therapy" reduced only systemic hypertension with no effect on glomerular hypertension and glomerular damage [25]. The same was described in uninephrectomised DOCA rats [26]. Raij and colleagues also described better effects of enalapril in relation to "triple therapy": enalapril reduced mesangial expansion and proteinuria [27]. Our study [8] as well as that of other authors [3, 25, 26] agree that ACE inhibitor captopril was better in comparison with hydralazine in slowing down glomerular sclerosis and mesangial expansion inspite of good regulation of blood pressure with both drugs. In ADR SHR, ACE inhibitors reduced proteinuria [6, 10, 20, 25], regulated systemic blood pressure (Fig. 1-a), decreased glomerular hypertension and glomerular sclerosis [7, 10, 25, 28, 29] which were also found in our experimental study (Fig. 1, Table 2). These were confirmed in clinical studies too: first, in patients with diabetic nephropathy [30] and later in patients with nondiabetic kidney diseases [6, 30-34]. In SHRs blood pressure increased from week 4 to 10, and after week 12 blood pressure was stabilized on a constantly higher level [36]. Our studied rats were 24 weeks old at the beginning of the study and they had stable hypertension in that period (Fig. 1-a). With the age in SHRs renal function aggravated very slowly, with little changes in glomeruli, higher in tubuloniterstitium [19] and the highest in blood vessels. SHRs developed glomerular changes very late inspite of hypertension, because glomeruli were protected with preglomerular vasoconstriction [37]. These mild changes described by various authors could be also seen in our study (Fig. 2, Table 2). Pathohistological changes in rat kidneys caused with anthracycline were first described by Stenberg and Phillips in 1967 [41]. Adriamycin (doxorubicin hydrochloride) induced syndrome nephroticum. Light microscopic study revealed no changes at the beginning of the study, but later (7-9 months) glomerular sclerosis, tubular dilatation and interstitial fibrosis developed which led to chronic renal failure [42, 43], (Fig. 3). Therapy with ACE inhibitor, captopril, in rats with adriamycin nephropathy lowered glomerular sclerosis [7, 12, 25, 28, 29], and mesangial expansion was very rare. In our earlier studies, captopril was found to decrease glomerular sclerosis in the early phase of adriamycin nephropathy in SHRs [8, 29], what was also confirmed in this study: captopril decreased glomerular and tubulointerstitial changes in weeks 6, 12 and 18 after second adriamycin injection, but had no effect on vascular changes (Fig. 4, Table 2). Favorable effects of captopril on tubulointerstiatial changes (especially interstitial) are very important because many authors have described recently better correlation between tubulointerstitial changes and CRF progression [17, 44-46], in comparison to glomerular changes and CRF. Besides, some authors have confirmed better effects of ACE inhibitors when they were given earlier before glomeruli were damaged [47]. CONCLUSION: In SHRs with ADR nephropathy treatment with captopril normalized systemic blood pressure, and slowed down CRF progression in their early stage. These functional changes correlate with significant slowing of glomerular and interstitial changes.