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-α) 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-α 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-α 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-α 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-α 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-α (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-α (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

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. [Projekat Ministarstva nauke Republike Srbije, br. 43012

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-β 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

Maternal smoking during pregnancy and socioeconomic factors as predictors of low birth weight in term pregnancies in Niš

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 characteristics of mothers who had children ≥ 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

Thyrocyte morphomethric analysis significance in differential diagnosis of thyroid carcinoma

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 μm2), and were significantly different from all other groups (p < 0.001). The mean nuclear area in FTC was 69.20 ± 27.31 μm2 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

Cytologic picture of 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

The red wine polyphenol, resveratrol improves hemodynamics, oxidative defence and aortal structure in essential and malignant hypertension

Resveratrol the polyphenolic antioxidant molecule has been proposed as an effective agent in the prevention of several pathological conditions, thus this study was aimed to evaluate its chronic effects on functional and structural remodeling of the aorta in spontaneously (SHR) and malignantly hypertensive rats (MHR). Resveratrol significantly decreased arterial pressure and improved regional hemodynamic parameters, bioavailability of nitric oxide, ameliorated morphological changes in the tunica media and smooth muscle of the aorta, preserved endothelium, reduced inflammation and apoptosis in both SHR and MHR, but excrete more beneficial effects on SHR. Resveratrol suppressed lipid peroxidation, significantly improved oxidative status and reduced levels of lipids. These results suggest the cardiovascular benefits of resveratrol dietary enrichment in both experimental models of hypertension

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