Institute for Medical Research

INTRODUCTION: Mesenchymal stem cells (MSCs) are a promising tool for regenerative medicine, but due to the heterogeneity of their populations, different sources and isolation techniques, the characteristics defining MSCs are inconsistent. OBJECTIVE: The aim of this study was to compare the characteristics of MSCs derived from two different human tissues: peripheral blood (PB-MSCs) and umbilical cord Wharton's Jelly (UC-MSCs). METHODS: The PB-MSC and UC-MSC were isolated by adherence to plastic after gradient-density separation or an explant culture method, respectively, and compared regarding their morphology, clonogenic efficiency, proliferating rates, immunophenotype and differentiation potential. RESULTS: MSCs derived from both sources exhibit similar morphology, proliferation capacity and multilineage (osteogenic, chondrogenic, adipogenic and myogenic) differentiation potential. Differences were observed in the clonogenic capacity and the immunophenotype, since UC-MSCs showed higher CFU-F (colony-forming units-fibroblastic) cloning efficiency, as well as higher embryonic markers (Na-nog, Sox2, SSEA4) expression. When additional surface antigens were analyzed by flow cytometry (CD44, CD90, CD105, CD33, CD34, CD45, CD11b, CD235a) or immunofluorescent labeling (vimentin, STRO-1 and alpha-smooth muscle actin), most appeared to have similar epitope profiles irrespective of MSC source. CONCLUSION: The results obtained demonstrated that both MSCs represent good alternative sources of adult MSCs that could be used in cell therapy applications.

Clary sage (Salvia sclarea L.) is native to Southern Europe and is cultivated worldwide. The essential oil of clary sage was analyzed as a potential antifungal agent. The main compounds in the oil were linalyl acetate (52.83%) and linalool (18.18%). Food poisoning agents, spoilage fungi, and plant and animal pathogens were among the tested fungal species. The microdilution method was used to establish minimal inhibitory concentrations (MIC) and minimal fungicidal concentrations (MFC). The commercial antimycotic bifonazole was used as a control. A concentration of 25 ?l/ml showed fungicidal activity against Aspergillus, Penicillium, and Fusarium species and Trichoderma viride. For the species Mucor mucedo and Aspergillus viride, the MFC was 15?l/ml; for Candida albicans, it was 10 ?l/ml, as in the case of bifonazole. Fungistatic and fungicidal activities of the oil against Cladosporium cladosporioides and Trichophyton menthagrophytes were recorded at concentrations of 2.5 ?l/ml and 5 ?l/ml. The most sensitive micromycetes were Cladosporium fulvum, Alternaria alternata, Phomopsis helianthi, and Phoma macdonaldii, where a concentration of 2.5 ?l/ml was lethal.

In many countries, including Croatia, there is no disease registry for osteoporosis and osteoporotic fractures. Epidemiological data show that the prevalence of osteoporosis is much higher in Asian people than in white European or North American populations, where the prevalence ranges between 10% and 15%. Epidemiological characteristics of osteoporosis in Croatia are similar to other European countries, though the incidence of osteoporotic fractures is somewhat higher. According to the annual report of registered diseases, the number of patients with osteoporosis and osteoporotic fractures increased during the last 10 years. In 2004, 0.70 % of all registered diseases in Croatia referred to osteoporosis and osteomalacia. The most important risk factors for osteoporosis are age, heredity, lifestyle, chronic diseases, hormonal abnormalities and physical characteristics of bone. Like in other chronic diseases, prevention measures are most important for disease control.

Abstract Erst in jüngster Zeit sind in der Entwicklung ionenselektiver (Glas‐, Festkörper‐, Flüssig‐) Membranelektrodensysteme zahlreiche Fortschritte erzielt worden. Die theoretischen Grundlagen und praktischen Voraussetzungen für derartige „Sensoren”︁ werden in diesem Beitrag skizziert, und es wird versucht, einen Überblick über ihre Anwendungsmöglichkeiten zu geben.

BACKGROUND: Toxoplasmosis has long been known as a major cause of perinatal morbidity. Acute infection in pregnancy may lead to fetal infection and subsequent fetal loss or birth of a manifestly or latently infected infant. However, it is a preventable disease. In Europe, significant variations have been shown to occur not only between countries but also within a given country indicating local variations in the influence of epidemiological factors contributing to infection. Thus, many European countries have implemented prevention programs in measure with the respective estimated risk of congenital toxoplasmosis. Since in view of its cost, a general screening-in-pregnancy program is at present not an option in Serbia & Montenegro, insight into the risk factors of particular local significance may therefore improve the quality of and the compliance with the hygienic and dietetic advice given to pregnant women as a preventive measure, as well as identify the particular subpopulations at an increased risk of infection, who may then be selectively screened. SUBJECTS AND METHODS: A retrospective study of risk factors for Toxoplasma gondii infection based on serological and epidemiological data (questionnaire) was performed in a series of 2936 women aged 15-49 years from throughout Serbia tested in our laboratory between 1988 and 1997. Inclusion criteria included availability of serological and epidemiological data (as specified below). Specific anti-Toxoplasma antibodies were detected by the reference Sabin-Feldman dye test as modified by Desmonts into the lysis test. The questionnaire included questions on age (stratified into five-year groups), degree of education (modalities: grade school, secondary or university level), and community of residence (urban/suburban), as well as on life-style habits pertaining to infection transmission risk factors: consumption of undercooked meat, exposure to soil, and exposure to cats (pet cat ownership). In addition, the year of entry into the study was taken as a variable per se. STATISTICS: The difference in the rates of infection between groups was evaluated by the chi-square test (chi 2). Logistic regression (univariant and multivariant approaches) was used to analyze the association between Toxoplasma infection and the specific demographic and epidemiological factors. RESULTS: The overall prevalence of infection was 69%, but with very wide variations both over time (decreasing from 86% in 1988 to 39% in 1997) and region (40-90%). The risk of infection increased from the south to the north (RR = 0.97, CI = 0.94-0.98). Within Belgrade, the risk was higher in urban than in suburban zones (RR = 0.79, CI = 0.64-0.93). The single infection transmission factor that was shown to be a predictor of infection in the whole study group was undercooked meat consumption (RR = 1.6, CI = 1.2-2.1), while exposure to soil was a predictor only in women aged 15-19 (RR = 10.3, CI = 2.7-38.6). Owing pet cats had no influence. While T. gondii infection was not associated with pathological pregnancies (p = 0.51) in the whole study population, it was significantly associated with pathological pregnancies in women exposed to both undercooked meat consumption (p = 0.009) and in those aged 15-19 in contact with soil (p = 0.022), as well as in women residing in highly urban communities (p = 0.048). CONCLUSION: The dramatic decrease in the prevalence of T. gondii infection over the nineties resulted in a rising proportion of women exposed to infection in pregnancy and consequently in an increased risk of congenital toxoplasmosis. Since the financial status of the country's health system does not allow for a general screening-in-pregnancy program, we propose, based on the above data, health education of all pregnant women in combination with serological testing of those exposed to predictors of infection as an epidemiologically sound and financially sustainable alternative.

INTRODUCTION: There is evidence that physical activity decreases the degree of atherosclerosis in the coronary arteries in persons who are physically active by modifying the chemistry and metabolism of lipoproteins. The effect of physical activity on the lipid status is achieved by affecting the enzymes of lipoprotein metabolism including the lipoprotein and liver lipase and transport protein of cholesterol esters. Epidemiological investigations on the effect of physical activity in prevention of cardiovascular diseases point to the fact that the persons who have a higher degree of cardio-respiratory endurance have a much lower mortality rate than those with a lower degree. The positive effect of physical activity on the cardiovascular system is reflected on the improved aerobic capacity, metabolic function, amplification of lipid profile, insulin sensitivity, immunological functions: it increases the perfusion of myocardium and the fibrinolytic activity, and reduces the adherence of thrombocytes due to increased synthesis of prostaglandin (PGI2), it also enhances the energy consumption, which is important in the maintenance of ideal bodyweight, prevention and treatment of obesity, and it has a positive effect on the control of stress. CONCLUSION: With respect to the effects on the lipid status, aerobic physical exercises like running, swimming, cycling, with the intensity of training of a medium (65% VO2) load, have a positive effect on lipid status. The best form of physical activity is the one in which the endurance and power are increased.

Low-grade systemic inflammation is at the base of the most chronic non-communicable diseases, which are reaching epidemic proportions worldwide. Key players in the regulation of inflammation are n-6 and n-3 polyunsaturated fatty acids (PUFAs), in particular arachidonic acid (n-6) and eicosapentaenoic acid (n-3). They are precursors of eicosanoids - signaling molecules involved in modulating the intensity and duration of inflammatory responses. Eicosanoids derived from n-6 PUFAs have proinflammatory actions, while those derived from n-3 PUFAs act anti-inflammatory. Therefore, dietary intake of n-6 and n-3 PUFAs, as well as their ratio, could markedly affect the pathogenesis and manifestation of many chronic diseases associated with low-grade inflammation. This review will focus on the relationship between dietary PUFAs and inflammation, with reference to PUFAs status in plasma phospholipids in Serbian population.

REGULATION OF INSULIN SECRETION: Beta cells are unique endocrine cells. They respond positively, in terms of insulin secretion, not only to changes in the extracellular glucose concentration, but also to activators of the phospholipase C (cholecystokinin or acetylcholine), and to activators of adenylate cyclase (glucagon, glucagon-like peptide-1, or gastric inhibitory polypeptide). Major messengers which mediate glucose action for insulin release are Ca2+, adenosine triphosphate (ATP) and diacylglycerol (DAG). MAJOR PATHWAYS OF INSULIN RELEASE STIMULATION: There are four major pathways involved in stimulation of insulin release. The first pathway is KATP channel-dependent pathway in which increased blood glucose concentrations and increased b-cell metabolism result in a change in intracellular ATP/ADP ratio. This is a contributory factor in closure of ATP-dependent K+ channels, depolarization of b-cell membrane, in increased voltage-dependent L-type Ca2+ channel activity. Increased Ca2+ influx results in increased intracellular Ca2+ and stimulated insulin release. KATP channel-independent pathway augments Ca(2+) -stimulated insulin secretion of KATP channel-dependent pathway. Major potentiation of release results from hormonal and peptidergic activation of receptors linked to adenylyl cyclase. Adenylyl cyclase activity is stimulated by hormones such as vasoactive intestinal peptide (VIP), glucagon-like peptide-1 (GLP-1), and so on. These hormones, acting via G protein, stimulate adenylyl cyclase, thus causing a rise in cyclic adenosine monophosphate (cAMP) and activation of protein kinase A (PKA). Increased activity of PKA results in potentiation of insulin secretion.

INTRODUCTION: Hyperlipoproteinemia is a key factor in development of atherosclerosis, whereas regression of atherosclerosis mostly depends on decreasing the plasma level of total and LDL-cholesterol. Many studies have reported the hypocholesterolemic effect of linolenic acid. TYPES OF POLYUNSATURATED FATTY ACIDS (PUFA): Linoleic and alpha-linolenic acids are essential fatty acids. The main sources of linoleic acid are vegetable seeds and of alpha-linolenic acid-green parts of plants. alpha-linolenic acid is converted to eicosapentaenoic and docosahexaenoic acid. Linoleic acid is converted into arachidonic acid competing with eicosapentaenoic acid in the starting point for synthesis of eicosanoids, which are strong regulators of cell functions and as such, very important in physiology and pathophysiology of cardiovascular system. Eicosanoids derived from eicosapentaneoic acid have different biological properties in regard to those derived from arachidonic acid, i.e. their global effects result in decreased vasoconstriction, platelet aggregation and leukocyte toxicity. ROLE AND SIGNIFICANT OF PUFA: The n-6 to n-3 ratio of polyunsaturated fatty acids in the food is very important, and an optimal ratio 4 to 1 in diet is a major issue. Traditional western diets present absolute or relative deficiency of n-3 polyunsaturated fatty acids, and a ratio 15-20 to 1. In our diet fish and fish oil are sources of eicosapentaenoic and docosahexaenoic acid. Refined and processed vegetable oils change the nature of polyunsaturated fatty acids and obtained derivates have atherogenic properties.

This work was a part of an initial study regarding the involvement of reactive nitrogen species (RNS) in paraquat (PQ) neurotoxicity. The nitrate concentration in the vulnerable regions of the brain (cortex, striatum and hippocampus) of Wistar rats was used as a measure of nitric oxide (NO) production or catabolism of the formed RNS. The tissue homogenates were deproteinized with acetonitrile and then centrifuged. Nitrate was measured in filtrated supernatants by simple and rapid isocratic ion-exchange high performance liquid chromatography with UV detection (IE-HPLC-UV) at 214 nm. The mobile phase (pH 8.5) consisted of borate buffer/gluconate concentrate, methanol, acetonitrile and deionized water (2:12:12:74, v/v/v/v), and the flow rate was 1.3 mL/min. Physiological nitrate levels (18.8 ? 6.1 nmol/mg of proteins), as well as a diverse range of nitrate concentrations could be determined with good precision (CV = 2.2 %) and accuracy (recovery of spiked samples was 99 ? 4%) in the brain tissue homogenates. Linearity was achieved in the range of nitrate from 0-80 ?M. The retention time of nitrate anion was 5.3 ? 0.3 min. .

To date, three types of dental stem cells have been isolated: Dental Pulp Stem Cells (DPSC), Stem Cells From Human Exfoliated Deciduous Teeth (SHED) and Immature Dental Pulp Stem Cells (IDPC). These dental stem cells are considered as mesenchymal stem cells. They reside within the perivascular niche of dental pulp. They are highly proliferative, clonogenic, multipotent and are similar to mesenchymal Bone Marrow Stem Cells (BMSC). Also, they have high plasticity and can be easy isolated. The expressions of the alkaline phosphatase gene, dentin matrix protein 1 and dentinsialophosphoprotein are verified in these cells. Analyses of gene expression patterns indicated several genes which encode extracellular matrix components, cell adhesion molecules, growth factors and transcription regulators, cell signaling, cell communication or cell metabolism. In both conditions, in vivo and in vitro, these cells have the ability to differentiate into odontoblasts, chondrocytes, osteoblasts, adipocytes, neurons, melanocytes, smooth and skeletal muscles and endothelial cells. In vivo, after implantation, they have shown potential to differentiate into dentin but also into tissues like bone, adipose or neural tissue. In general, DPSCs are considered to have antiinflammatory and immunomodulatory abilities. After being grafted into allogenic tissues these cells are ableto induce immunological tolerance. Immunosuppressive effect is shown through the ability to inhibit proliferation of T lymphocytes. Dental pulp stem cells open new perspectives in therapeutic use not only in dentin regeneration, periodontal tissues and skeletoarticular, tissues of craniofacial region but also in treatment of neurotrauma, autoimmune diseases, myocardial infarction, muscular dystrophy and connective tissue damages.

Green areas at four localities in the Belgrade region (Ada Ciganlija, Kosutnjak, Miljakovac forest, and Mt. Avala) were investigated in 2004. The aim of the research was to clarify the faunistic composition, relative abundance, and population dynamics of ticks, as well as the seasonal distribution of Borrelia burgdorferi sensu lato (sl) in Ixodes ricinus. Two species of ticks were detected: Ixodes ricinus and Dermacentor reticulates. Relative abundance analysis revealed that the species Ixodes ricinus was predominant (97.41 %). Out of 942 Ixodes ricinus ticks, 188 (19.96 %) were infected with Borrelia burgdorferi sl. The infection rate of adults by localities ranged from 19.16% to 30.99% (Mt. Avala and Ada Ciganlija, respectively).

INTRODUCTION: Growth is regulated by the interaction of environmental signals with endogenous neuroendocrine responses to the genetic programs that determine the body plan. The insulin-like growth factors (IGFs) are integral components of multiple systems controlling both growth and metabolism. THE IGF SYSTEM: The IGF system is thouht to be more complex than other endocrine systems, as genes for six IGF-binding proteins (IGFBPs) have been identified so far. The IGFs play a critical role in both cell cycle control and apoptosis, two functions involved in regulation of tumorigenesis. Insulin-like growth factor-I (IGF-I) is essential for normal growth. Confirmation of the significance of IGF-I in human physiology was obtained by the discovery of a patient with intrauterine growth retardation and postnatal growth failure associated with a mutation in the IGF-I gene. STAGES OF EVOLUTION OF THE SOMATOMEDIN HYPOTHESIS: The original somatomedin hypothesis postulated that somatic growth was regulated by growth hormone's (GH's) stimulation of hepatic IGF-I production, with IGF-I acting in an endocrine fashion to promote growth. The dual effector theory proposed an alternative view, involving direct effects by GH on peripheral tissues not mediated by IGF-I and GH-stimulated local IGF-I production for autocrine/paracrine action. It is now clear that GH stimulates the formation of ternary IGF binding complex, which stabilizes IGF-I in the serum.

One of the most important breakthroughs in the understanding of bone biology was the identification of the role of cytokines in bone remodelling including the alveolar bone exposed to the effect of mechanical forces during orthodontic treatment. Since bone remodelling is associated, in its early phase, with inflammation of the surrounding tissue, the hypothesis has been suggested on the role of proinflammatory cytokines in the process of bone remodelling, primarily IL-1beta, IL-6 and TNFalpha. These cytokines function as response mediators in the acute phase of inflammation, as well as in the processes of metabolism, and stimulation of resorption and inhibition of bone formation. Mostly uninvestigated, the dynamics of concurrent changes of these three cytokines during the early phase of orthodontic teeth movement in children and adults was the subject of our investigation presented in this article on the current knowledge on the role of cytokines in this process.

Stimulation of glutamate receptors induces neuronal nitric oxide (NO) release, which in turn modulates glutamate transmission. The involvement of ionotropic glutamate NMDA and AMPA/kainate receptors in induction of NO production in the rat brain was examined after injection of kainate, a non-NMDA receptor agonist; kainate plus 6-cyano- 7-nitroquinoxaline-2,3-dione (CNQX), a selective AMPA/kainate receptor antagonist; or kainate plus 2-amino-5-phosphonopentanoic acid (APV), a selective NMDA receptor antagonist. Competitive glutamate receptor antagonists were injected with kainate unilaterally into the CA3 region of the rat hippocampus. The accumulation of nitrite, the stable metabolite of NO, was measured by the Griess reaction at different times (5 min, 15 min, 2 h, 48 h, and 7 days) in hippocampus, forebrain cortex, striatum, and cerebellum homogenates. The used glutamate antagonists APV and CNQX both provided sufficient neuroprotection in the sense of reducing nitrite concentrations, but with different mechanisms and time dynamics. Our findings suggest that NMDA and AMPA/kainate receptors are differentially involved in nitric oxide production. <br><br><font color="red"><b> This article has been retracted. Link to the retraction <u><a href="http://dx.doi.org/10.2298/ABS150319031E">10.2298/ABS150319031E</a><u></b></font>

Helicobacter pylori (H. pylori) colonizes the gastric mucosa of a half of the mankind. Duodenal ulcer is found in 15-25%, gastric ulcer in 13%, while gastric adenocarcinoma develops in 1% of all infected individuals. Pathogenesis of H. pylori infection is related to the virulence factors of the bacterium, environmental (dietary habits, hygiene, stress) and host factors (age, sex, blood type). Colonization of the gastric mucosa is related to the motility of the bacterium, presence of lipopolysaccharide (LPS) and various bacterial enzymes. Gastric mucosal injury is the result of H. pylori LPS, vacuolization cytotoxin (vacA), cytotoxin associated protein (cagA), heat shock proteins and factors responsible for neutrophil chemotaxis and activity. H. pylori colonizes the gastric mucosa and zones of ectopic gastric epithelium. H. pylori infection is transmitted via oral-oral, fecal-oral and iatrogenic way (during endoscopy). Higher prevalence of the infection is associated with lower socioeconomic level, lack of drinking water, and living in a community. Acute H. pylori gastritis is superficial pangastritis progressing into the chronic phase after 7-10 days. Gastric mucosal atrophy and intestinal metaplasia can develop during the course of H. pylori infection. Clearly defined factors that influence the outcome of H. pylori infection include bacterial strain, distribution of gastritis, acid secretion and gastric mucosal atrophy.

Automatic text analysis is a methodological approach in the analysis of individual differences in verbal behaviour. It enables extraction of statistically manipulable information about intensity and/or frequency of thematic and stylistic characteristics of verbal output. LIWC (Linguistic Inquiry and Word Count), one of the widely used software solutions for automatic text analysis, performs analyses by matching word stems from incorporated software dictionary with those from text input. It provides information about the percentage of each of the predefined categories in the analyzed text. Research suggests that data obtained by automatic text analysis have potential in explaining the relationship between implicit and explicit measures, independently of the object of measurement (attitudes, pathological potential, assessment of basic personality traits etc.). The topic of this paper is the development of the Serbian LIWC dictionary. Development of the dictionary was performed in four phases: translation of English LIWC dictionary, forming lemmas, classification of word stems according to absolute consensus among four independent raters (where word stems could be categorized in more than one category, depending on the context), and revision of the content of categories and creation of final set of word stems. The final version of the LIWCser dictionary contains 12103 word stems classified into 65 categories (linguistic, psychological and personal concerns). Only four word stems (0.03%) were classified into eight categories, 22 (0.2%) into seven, 147 (1.2%) into six, and 568 (4.7%) into five. 1531 (12.6%) word stems were classified into four categories, 2913 (24.1%) into three, 4800 (39.7%) into two, while 2188 (17.5%) word stems were classified into only one category. Development of the LIWCser dictionary allows researchers to collect and analyze data on verbal behaviour and to study the relationship between implicit and explicit measures in different fields of psychology.

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.

BACKGROUND/AIM: The beneficial effects of medicinal plants are well-known from the ancient times. However, expansion of phytopharmacy and phytotherapy occured during the last decades. Medicinal plants can absorb environmental contaminants from the ground and consequently may cause harmful effects on human health. Quality control usually comprises standard methodology which includes macroscopic identification and examination of active ingredients. Additionally, there is a permanent need to control the level of pollutants in herbs, with a particular attention to the level of toxic metals. In this study we estimated the level of contamination by determining the content of cadmium (Cd) in the herbs of Hypericum perforatum and Thymus serpyllum collected from the different localities of the mountains Rtanj and Ozren. METHODS: Herbs of investigated plants were collected during July 2005 from various localities of Rtanj and Ozren mountains. After drying, homogenization and mineralization, Cd content was determined by atomic absorption spectrophotometry. RESULTS: The obtained results show that Cd content varies significantly in Hypericum perforatum samples collected. The lowest Cd level was found in samples from the one of Rtanj localities (0.25 mg Cd/kg), while the highest was observed in Hypericum peforatum from Ozren locality (1.24 mg/kg). Levels of Cd in the three of four investigated localities were higher than proposed by WHO (0.3 mg/kg dried herb material). In all investigated samples of Thymus serpyllum herbs Cd levels were below the limit of detection of analytical method (0.2 mg Cd/kg dried materials). CONCLUSION: This work contributes to the issue of Cd content in Hypericum perforatum and Thymus serpyllum grown in localities of Rtanj and Ozren, and implies the importance for systemic control of Cd content in Hypericum species in order to provide safety of their preparations. Furthermore, regarding Cd toxicity, maximal permissible level of Cd in plant material should be evaluated and established concerning national legislative frame.

INTRODUCTION: Our study was aimed at determining whether the polymorphism of genes for different components of the renin-angiotensin-aldosterone system could influence the renal hemodynamic response to losartan treatment. MATERIAL AND METHOD: The study included 35 patients with type I diabetes mellitus and persistent albuminuria, genotyped for the 1166 A/C polymorphism gene for the angiotensin II type 1 receptor and I/D polymorphism of the angiotensin-converting enzyme gene. The participants were divided into groups according to the combinations of A or C allele: AA, AC, CC; and according to the combinations of I or D allele: II, ID and DD genotype. The patients received losartan therapy for 12 weeks. The renal hemodynamic measurements were determined at baseline and after the examination period. RESULTS: Losartan therapy significantly reduced the filtration fraction from the baseline by 0.018 +/- 0.024 (p = 0.012) only in the AC genotype. The glomerular filtration rate remained unchanged in all genotype groups. A significant increase in the effective renal plasma flow was obtained only in AC genotype (544 +/- 88 vs 575 +/- 90 ml/min; p = 0.02), while significant reductions in the renal vascular resistance were found in AA group (115 +/- 25 vs 95 +/- 21 mmHg x l(-1) x min(-1), p = 0.001) and in AC group (118 +/- 30 vs 101 +/- 28 mmHg x l(-1) x min(-1); p = 0.001). A significant reduction of the glomerular filtration rate by 8 +/- 10 ml/min was obtained only in the DD genotype (p = 0.016), and only the DD genotype achieved a significant reduction of the filtration fraction by 0.019 +/- 0,022 (p = 0.008). The most pronounced increase of the effective renal plasma flow was found only in the ID genotype (536 +/- 75 vs 591 +/- 63 ml/min; p = 0.01). The reduction of the renal vascular resistance was independent of ACE gene polymorphism. CONCLUSION: Our study shows that individual renal vascular response to losartan treatment in diabetic patients with persistent albuminuria, could be influenced by genetic polymorphisms.