Resveratrol Protects Cardiac Tissue in Experimental Malignant Hypertension Due to Antioxidant, Anti-Inflammatory, and Anti-Apoptotic Properties.

Hypertension is one of the most prevalent and powerful contributors of cardiovascular diseases. Malignant hypertension is a relatively rare but extremely severe form of hypertension accompanied with heart, brain, and renal impairment. Resveratrol, a recently described grape-derived, polyphenolic antioxidant molecule, has been proposed as an effective agent in the prevention of cardiovascular diseases. This study was designed to examine chronic resveratrol administration on blood pressure, oxidative stress, and inflammation, with special emphasis on cardiac structure and function in two models of experimental hypertension. The experiments were performed in spontaneously (SHRs) and malignantly hypertensive rats (MHRs). The chronic administration of resveratrol significantly decreased blood pressure in both spontaneously and malignant hypertensive animals. The resveratrol treatment ameliorated morphological changes in the heart tissue. The immunohistochemistry of the heart tissue after resveratrol treatment showed that both TGF-ß and Bax were not present in the myocytes of SHRs and were present mainly in the myocytes of MHRs. Resveratrol suppressed lipid peroxidation and significantly improved oxidative status and release of NO. These results suggest that resveratrol prevents hypertrophic and apoptotic consequences induced by high blood pressure with more pronounced effects in malignant hypertension.

Smoking habits, knowledge about and attitudes toward smoking among employees in health institutions in Serbia.

BACKGROUND/AIM: According to the number of active smokers, Serbia occupies a high position in Europe, as well as worldwide. More than 47% of adults are smokers according to WHO data, and 33.6% according to the National Health Survey Serbia in 2006. Smoking physicians are setting a bad example to patients, they are uncritical to this habit, rarely ask patients whether they smoke and rarely advise them not to smoke. These facts contribute to the battle for reducing the number of medical workers who smoke, as well as the number of smokers among general population. The aim of the study was to determine the smoking behavior, knowledge and attitudes and cessation advice given to patients by healthcare professionals in Serbia. METHODS: A stratified random cluster sample of 1,383 participants included all types of health institutions in Serbia excluding Kosovo. The self administrated questionnaire was used to collect data about smoking habits, knowledge, attitudes and cessation advice to patients given by health professionals in Serbia. RESULTS: Out of 1,383 participants, 45.60% were smokers, of whom 34.13% were physicians and 51.87% nurses. There were 46.4% male and 45.4% female smokers. The differences in agreement with the statements related to the responsibilities of health care professionals and smoking policy are significant between the "ever" and "never" smokers, and also between physicians and nurses. Twenty-five percent of nurses and 22% of doctors claimed they had received formal training. However, only 35.7% of the healthcare professionals felt very prepared to counsel patients, while 52.7% felt somewhat prepared and 11.6% were not prepared at all. CONCLUSIONS: According to the result of this survey, there are needs for more aggressive nationwide non-smoking campaigns for physicians and medical students. Experiences from countries where physicians smoke less and more effectively carry out smoking cessation practices need to be shared with Serbian physicians in order to improve their smoking behavior and smoking cessation practices.

Efficacy of amifostine in protection against doxorubicin-induced acute cardiotoxic effects in rats.

BACKGROUND/AIM: Amifostine (AMI) is a broad-spectrum cytoprotector which protects against variety of radio- and chemotherapy-related toxicities without decreasing their antitumor action. The aim of the study was to investigate the potential protective effects of AMI against acute cardiotoxic effects of doxorubicin (DOX) in male Wistar rats. METHODS: AMI (300 mg/kg ip) was given 30 min before DOX (6 mg/kg and 10mg/kg b.w., iv). The evaluation of DOX-induced cardiotoxic effects, as well as cardioprotective efficacy of AMI was performed 48 h after their administration by determining serum activities of enzymes known to be markers of cardiac damage (creatine kinase - CK, aspartate aminotransferase - AST, lactate dehydrogenase - LDH, and its isoenzyme alpha-hydroxybutirate dehydrogenase - alpha - HBDH), as well as the histopathological and ultrastructural analysis of the heart tissue. RESULTS: AMI successfully prevented a significant increase in serum activity of CK, AST, LDH and alpha-HBDH in animals treated with DOX in the dose of 6 mg/kg (121.14 +/- 18.37 vs 167.70 +/- 44.24; 771.42 +/- 161.99 vs 1057.00 +/- 300.00; 3230.00 +/- 1031.73 vs 4243.10 +/- 904.06; 202.57 +/- 42.46 vs 294.90 +/- 80.20 UI/l, respectively), and ameliorated DOX-induced structural damage of the rat myocardium. Pretreatment with AMI in rats given 10 mg/kg DOX reduced the cardiac damage score (CDS) from 2.62 +/- 0.51 to 1.62 +/- 0.51, i.e. to the CDS value obtained with the lower dose of DOX (6 mg/kg). The ultrastructural analysis of the rat myocardium showed that AMI successfully protected the sarcolemma of cardiomyocytes and reduced mitochondria damage induced by DOX given in the dose of 6 mg/kg. Besides, capillaries were less morphologically changed and apoptosis of endothelial cells was extremely rare in AMI-protected animals. AMI itself did not cause any prominent changes in the examined parameters in comparison with the control rats. CONCLUSION: AMI provided a significant protection against DOX-induced acute cardiotoxic effects in rats. This finding implies its potential to be a successful cardioprotector in patients treated with DOX due to malignant diseases.

[Thyrocyte morphomethric analysis significance in differential diagnosis of thyroid carcinoma].

UNLABELLED: BACKGRAUND/AIM: Conventional cytomorphology of thyroid cell aspirates is limited in preoperative differential diagnosis of follicular adenomas (FA) and hyperplastic adenomatoid nodular goiters from well differentiated thyroid follicular carcinoma (FTC) and follicular variant of thyroid papillary carcinoma (PTC). This is the reason of inaccurate presurgical differential diagnosis and in the same cases of inadequate operative managament. The aim of the study was to evaluate the role of quantitative estimation of thyreocite nuclear features (cariomorphometry) in thyroid aspirated smears in preoperative differential diagnosis of benign from malignant thyroid lesions. METHODS: A total of 48 patients with thyroid nodular disease underwent fine needle aspiration biopsy for cytomorphology, cariomorphometric analysis of the aspirates, and histopathologic explorations conducted fully postoperatively. On the basis of cytomorphology classification the patients were divided into three groups: benign (B), n = 8; malignant (M), n = 15, and suspicious for malignancy (S), n = 25. Using a microscope connected to a computerized video system, mean nuclear area, the nuclear area coefficient of variation (NACV) and anisocariosis ratio were measured and calculated. RESULTS: In all the 15 patients with cytologically malignant results the diagnosis of PTC was confirmed histopathologically. All cytologically benign lesions were confirmed histopathologically. Thyroid carcinoma was found in 15 out of 25 patients with suspicious lesions. The highest mean values of nuclear area were in the PCT (90.74 +/- 26.71 microm2), and were significantly different from all other groups (p < 0.001). The mean nuclear area in FTC was 69.20 +/- 27.31 microm2 and was significantly higher than in the benign adenomatous group (p < 0.01). There was no significant difference in mean nuclear area between FTC and FA, but there was a significant difference in NACV between these two groups (FTC: 39.46% vs FA: 23.42%,p < 0.001). In 27 out of 30 patients with thyroid carcinoma higher values of NACV than 18% were found. CONCLUSION: Preoperatively cariomorphometry is a useful method in differential diagnosis of thyroid carcinoma from benign lesions, as a complementary method to convencional cytodiagnostics. The NACV showed highest sensitivity as a parameter of malignant thyroid cell transformation.

Skin vascularisation field by the ascending branch of the peroneal artery ramus perforans.

BACKGROUND/AIM: Soft tissue defects in the distal third of the lower leg are persistent and constitute a major problem in the reconstructive surgery. This study presents an analysis of the anatomical vascularization filed of ascending branch of the peroneal artery ramus perforans (PARS). The aim of this study was to assess reliability of the distal flap on the antero-lateral aspect of a lower leg distal third. METHODS: Direct gentiana violet injection into the interosseal perforator of ten fresh cadaveric lower legs with subsequent corrosion acrylic preparation was performed to reveal vascularization filed of the ascending branch of the PARP. Height, length, diameter and communication of perforating branch and its subsequent smaller ascending and descending branches were determined. The CAMIA software was used. RESULTS: Our results show that the PARP is always present. Its origin from the peroneal artery is at the medial height of 66 mm when measured from the inferior border of the lateral malleolus. Medium length of ramus perforans is 51.7mm. After transition through the interosseous membrane, ramus perforans divides into ascending and descending branches. The diameter proximal to the level of bifurcation is 1.37 mm (variation 1.0-1.8 mm), and the diameter of the ascending branch distal to the level of bifurcation is 1 mm. Using CAMIA software, the medium length, width and area of the vascularization filed labeled with gentian violet were calculated to be 164 mm (variation 125-210 mm), 66 mm (57-77 mm), and 10,305 mm2 (6,385 mm2-14,341 mm2), respectively. CONCLUSION: Our results support the use of fasciocutaneous distal flap, vascularized by the ascending branch of the PARP for reconstruction of soft tissue defects in the distal third of the lower limb, malleolar regions and dorsum.

[Evaluation of morphological changes of the liver caused by heroin abuse in forensic practice].

BACKGROUND/AIM: A study of morphological lesions in the liver of heroin addicts enables a precise overview of the type and degree of the liver damages caused by intravenous (i.v.) heroin abuse, additive effects of viral infections and alcohol consumption, as well as whether the expressiveness of these lesions depends on the duration of the time period of heroin application. The aim of the study was to investigate histopathological, ultrastructural and morphometric features of the liver of heroin addicts in forensic samples of the liver. METHODS: The study involved the autopsy conducted on 40 bodies of i.v. heroin addicts and 10 control autopsies. The investigated group consisted of liver samples of 36 male subjects and 4 female subjects aged 35-40 years and the control group of 8 male and 2 female cadaveric bodies aged 15-35 years. The liver tissue samples were prepared for light microscopy. Sections of the tissue paraffin blocks 5 micro thick were stained using classical Hematoxylin and Eosin method (H&E), as well as PAS Van Gieson, Gomori, and Congo Red techniques. For investigation purposes of ultrastructural changes, liver tissue was fixed in glutaraldehyde and molded with epon. The analysis was performed using the method of transmission electron microscopy. Morphometric investigation of the liver sinusoidal macrophages was performed by using the M42 test system. RESULTS: In the investigated group of i.v. heroin addicts, the liver autopsy samples showed degenerative vesicular and fat changes, chronic hepatitis, cirrhosis, sedimentation of pathologic protein amyloidosis, dysplastic changes, reduction in the amount of glycogen in hepatocytes, as well as the change in the number of Kupfer and endothelial cells. The established changes correlated with the duration of i.v. heroin abuse, whereas sinusoidal macrophages were activated in cases with active hepatitis, and no significant change in their number was found in hepatocytes with alcohol-related fatty changes. CONCLUSION. The study showed that the most present change in the hepatocytes of drug addicts was vesicular degeneration, and it is the only direct consequence of the effect of heroin. Other morphological changes were present due to viral infections and they correlated with the duration of narcotic abuse. The finding of dysplastic changes in this susceptible population of young people is particularly significant. The forensic significance of the established changes in the liver tissue is in the possibility of their practical application for determination of the immediate cause of death of i.v. heroin addicts, as well as the differential diagnosis of not only heroin, but also alcohol, sedative and other substances abuse, and all that on the basis of morphological damages of the liver.

[Maternal smoking during pregnancy and socioeconomic factors as predictors of low birth weight in term pregnancies in Nis].

BACKGROUND/AIM: Low birth weight (LBW) is a result of preterm birth or intrauterine growth retardation, and in both cases is the strongest single factor associated with perinatal and neonatal mortality. It is considered that socioeconomic factors, as well as mothers bad habits, play the most significant role in the development of LBW, which explains notable number of researches focused on this particular problem. The aim of this study was to characterize socioeconomic factors, as well as smoking habits of the mothers, and their connection with LBW. METHODS: The questionnaire was carried out among mothers of 2 years old children (n = 956), born after 37 gestational weeks. The characteristics of mothers who had children with LBW, defined as < 2,500 g, (n = 50), were matched with the characteristices of mothers who had children > or = 2,500 g, (n = 906). For defining risk factors, and protective factors as well, we used univariant and multivariant logistic modeles. RESULTS: As significant risk factors for LBW in an univariant model we had education level of the mothers, smoking during pregnancy, smoking before pregnancy, the number of daily cigarettes, the number of cigarettes used during pregnancy, paternal earnings and socioeconomic factors. In a multivariant model the most significant factors were socioeconomic factors, education level of the mothers, paternal earnings and mothers smoking during pregnancy. CONCLUSION: Smoking during pregnancy and socioeconomic factors have great influence on LBW. Future studies should be carried out in different social groups, with the intention to define their influence on LBW and reproduction, as well. This should be the proper way of adequate health breeding planning for giving up smoking, the prevention of bad habits and melioration of mothers and children health, as the most vulnerable population.

[Cytologic features of ameloblastoma]. / Citoloska slika ameloblastoma.

Ameloblastoma is a rare tumor of the jaw arising from odontogenic epithelium. There are sparse reports in the literature concerning cytologic features of this tumor. This paper presents two cases of ameloblastoma, diagnosed by imprint cytology and confirmed histopathologically. The imprints were hypercellular, with single cells and the groups of basaloid and polygonal squamous cells with huge vacuoles in cytoplasm. Stellate and fusiform cells were found in the background of the preparation. These morphologic parameters were sufficient for the cytologic diagnosis of ameloblastoma.

Lupus nephritis: histopathologic features, classification and histologic scoring in renal biopsy.

Lupus nephritis (LN) is one of the most significant manifestations in systemic lupus erythematosus (SLE), although in some cases there is no direct correlation among clinical, serologic and histologic findings. Therefore, renal biopsy and histopathologic classification by the activity and chronicity assessment of LN are considered necessary before the initiation of the treatment. In this paper 311 renal biopsies in patients with LN were analyzed and classified according to the WHO Classification published in 1974. Renal biopsy specimens were routinely processed for standard analysis by light microscopy (LM), immunofluorescent (IF), and electron microscopy (EM). Biopsy findings were compared with the common clinical symptoms. It is important to recognize that histopathologic lesions in the analyzed tissue might precede the symptoms and clinical manifestations, and therefore are an important factor in the appropriate therapeutic approach. Histopathological assessment of the class of LN represents an important predictor in most patients because the lesions of LN class I and class II most often have no progression and consequently do not require an aggressive treatment. Diffuse proliferative glomerulonephritis (class IV) as a severe form of renal lesion requires the application of high doses of corticosteroids and cytotoxic medicaments. The established quantitive indexes have significant predictive value. Activity index (AI) evaluates the presence of fresh inflammatory and potentially reversible lesions and chronicity index (CI) evaluate the presence of irreversible glomerular lesions such as sclerosis, tubular atrophy, interstitial fibrosis, etc. High AI and low CI, particularly in the class IV of LN, suggest the necessity of an aggressive treatment of the inflammatory process aiming to preserve the renal function. High CI and low AI characterize irreversible sclerosing lesions, where an aggressive treatment is considered unnecessary. Our experience suggests the necessity of introducing the new morphologic elements into the activity and chronicity scoring system which would correlate better the clinical symptoms. This should provide higher reliability of scoring, since it evaluates important elements in clinical-morphologic diagnosis of LN.

Production of TNF-alpha by skin explants of dinitrochlorobenzene-challenged ears in rats: a model for the evaluation of contact hypersensitivity.

BACKGROUND: Contact hypersensitivity (CHS) is a local inflammatory response of the skin following challenge of hapten-sensitized animals. It is the consequence of cell infiltration of derm and the release of inflammation mediators, among which Tumor necrosis factor-alpha (TNF-alpha) is one of the most important factors. The intensity of the inflammation could be quantified by ear swelling which is the classical manifestation of the reaction. This study was testing the working hypothesis that levels of TNF-alpha in skin organ culture medium should correlate with the intensity of CHS reaction measured in vivo by ear swelling assay, and with the density of dermal infiltrate in ear skin samples. In order to test the working hypothesis, the intensity of inflammatory reaction following challenge was evaluated by classical measurements of ear swelling, by the determination of TNF-alpha levels in culture fluids of ear skin following epicutaneous application of dinitrochlorobenzene (DNCB) into the ears of sensitized animals. METHODS: Animal model of CHS reaction to DNCB in Albino Oxford rats was used as described. Ear swelling was quantified in percentage terms as the difference in thickness between the challenged and nontreated ears of the same animal. Dermal infiltrate density in histopathologically analyzed samples of ear skin was evaluated by computer-assisted image analysis. Ear skin samples were cultured in standard medium for 24 h, and TNF-alpha concentration in the conditioned medium was subsequently determined with ELISA test. RESULTS: Dose-dependent increase in the density of the dermal infiltrate and in TNF-alpha in CM were noted following the application of 0.65%, 1.3% and 2.6% of DNCB to the ears of previously sensitized rats. The correlation between ear swelling and the levels of TNF-alpha (r = 0.933, p < 0.001) in CM, and between ear swelling and dermal infiltrate density (r = 0.916, p < 0.001) was found. Correlation was also found between the density of the dermal infiltrate and the levels of TNF-alpha (r = 0.865, p < 0.001). CONCLUSION: Presented data suggested that skin-organ culture system and the quantification of inflammatory mediators might be used for the evaluation of contact hypersensitivity reaction and its intensity.

[Diagnosis of the Mikulicz-Sjogren syndrome using biopsy of the minor salivary glands]. / Dijagnostika sindroma Mikulicz-Sjögren biopsijom malih pljuvacnih zlezda.

BACKGROUND: Sjögren's syndrome (SS) represents autoimmune disease characterized by chronic inflammation, destruction and insufficiency of exocrine glands, particularly salivary and lacrimal glands, accompanied by dryness of mouths and eyes. Diagnostic work-up involves clinical laboratory tests, radiography, scintigraphy and bioptic histopathological examination. Examination of small salivary glands in the biopsy of the lower lip represents a "golden standard" of diagnosis of SS, concerning the fact that the growth and the disfunction of salivary and lacrimal glands occurs in different pathologic states. METHODS: Resected specimens of the lower lip were obtained from 47 patients with clinical diagnosis of SS. After standard histopathological treatment, slices were hematoxylin and eosin stained. Immunohistochemistry against smooth muscle actin was performed using LSAB+ method (AHSMA-M7558, DAKO 1:50). On the basis of generally accepted histopathological diagnostic criteria the results were categorized as: findings suspicious for SS; findings compatible with the diagnosis of SS (mild, moderate and high degree of inflammation); nonspecific inflammatory reaction and nonrepresentative biopsy samples. RESULTS: Diagnosis of SS was confirmed in 32% of cases. In 2% of cases findings were suspected for SS, in 36% of cases findings were compatible with the diagnosis of nonspecific inflammation, and in 30% of cases material was not representative. CONCLUSIONS: By the biopsy of salivary glands of the lower lip the diagnosis of SS was confirmed in 50-60% of cases. Upon the precise diagnostic criteria it was also possible to determine the intensity of inflammation and tissue destruction in SS and identify other pathological conditions, which justified the biopsy. Surgical technique had to be adequate in order to obtain representative number of small salivary glands. In the presented material 30% of specimens were nonrepresentative which was very high percentage compared with literature data. This was most probably the reason why the diagnosis of SS was confirmed in only 32% of cases, i.e., in every third patient.