THE EFFECTS OF ENDOCRINE DISRUPTORS ON FEMALE GONADAL AXIS: AN UPDATE.

Endocrine disruptors (EDs) are considered to have an impact on the function of reproductive axis at different levels as well on reproductive organs in both sexes. Complexity of female reproductive system influenced with various stressors including EDs lead to morphological and functional alterations. This is resulting in modulation of neuroendocrine regulation with consequent developmental irregularities and derangements, causative infertility, endometriosis as well as premature ovarian insufficiency or polycystic ovary syndrome. A number of experimental clues was obtained on female animal models using various EDs such as synthetic estrogens and phytoestrogens, neurotransmitters, pesticides or various chemicals. These substances lead towards consequent derangement of the neuroendocrine control of reproduction from early phases of reproductive development towards different phases of adult reproductive period. This text will address some novel insights into the effects of EDs on neuroendocrine regulation of gonadal axis, effects on ovaries as well on endometrium during implantation period.

Biochemical response of hybrid black poplar tissue culture (Populus × canadensis) on water stress.

In this study, poplar tissue culture (hybrid black poplar, M1 genotype) was subjected to water stress influenced by polyethyleneglycol 6000 (100 and 200 mOsm PEG 6000). The aim of the research was to investigate the biochemical response of poplar tissue culture on water deficit regime. Antioxidant status was analyzed including antioxidant enzymes, superoxide-dismutase (SOD), catalase (CAT), guiacol-peroxidase (GPx), glutathione-peroxidase (GSH-Px), glutathione-reductase, reduced glutathione, total phenol content, Ferric reducing antioxidant power and DPPH radical antioxidant power. Polyphenol oxidase and phenylalanine-ammonium-lyase were determined as enzymatic markers of polyphenol metabolism. Among oxidative stress parameters lipid peroxidation, carbonyl-proteins, hydrogen-peroxide, reactive oxygen species, nitric-oxide and peroxynitrite were determined. Proline, proline-dehydrogenase and glycinebetaine were measured also as parameters of water stress. Cell viability is finally determined as a biological indicator of osmotic stress. It was found that water stress induced reactive oxygen and nitrogen species and lipid peroxidation in leaves of hybrid black poplar and reduced cell viability. Antioxidant enzymes including SOD, GPx, CAT and GSH-Px were induced but total phenol content and antioxidant capacity were reduced by PEG 6000 mediated osmotic stress. The highest biochemical response and adaptive reaction was the increase of proline and GB especially by 200 mOsm PEG. While long term molecular analysis will be necessary to fully address the poplar potentials for water stress adaptation, our results on hybrid black poplar suggest that glycine-betaine, proline and PDH enzyme might be the most important markers of poplar on water stress and that future efforts should be focused on these markers and strategies to enhance their concentration in poplar.

[NEPHROTOXIC DRUGS].

Renal tissue is sensitive to the effect of potentially nephrotoxic drugs and other substances that are available over-the-counter or can be purchased at healthy food stores or elsewhere, and harmful substances from the environment. The harmful effects of these substances lead to the development of recognizable clinical syndromes, including acute or chronic renal failure, tubulopathy, and proteinuria. Risk factors that influence the development of kidney disease induced by drugs are divided into those related to patient characteristics, drug characteristics, and renal function. Drugs that commonly exhibit nephrotoxic effects are analgesics, antimicrobials, chemotherapeutics, contrast agents, immunosuppressants, herbal preparations and substances containing heavy metals. Family physician must carefully observe their patients, nurturing individual approach to drug selection and determining the dose. Renal function can quickly return to normal if the damage is recognized on time. Recent research yields insights into the identification of new biomarkers that will contribute to early detection of drug induced kidney damage.

[DIET CHARACTERISTICS IN PATIENTS WITH CHRONIC KIDNEY DISEASE].

Because of the increasing number of patients, chronic kidney disease (CKD) has become a significant public health problem. As kidney function decreases, it is necessary to introduce certain dietary modifications. The aim was to investigate what is the appropriate approach to diet of CKD patients, which could contribute to slowing down progression of the disease. Dietary recommendations are individual for each patient, but also vary in the same patient depending on the stage of disease progression because special attention must be paid to appropriate intake of macronutrients (protein, carbohydrates and fats), micronutrients (sodium, potassium, calcium, phosphorus, zinc, selenium, various vitamins), and water. In newly diagnosed patients, it is necessary to assess their nutritional status and energy requirements. It has been shown that protein-energy malnutrition, muscle loss and cachexia are strong predictors of mortality in CKD. Comparing different dietary approaches in everyday life of patients suffering from CKD, it was found that the most effective diet is Mediterranean food style. Studies confirm that Mediterranean diet has a preventive effect on renal function and reduces progression of the disease. Preventive measures, correct identification and early intervention can increase survival of patients and improve their quality of life. Mediterranean diet tailored to individual stages of CKD has been confirmed as the best choice in CKD patients.

[PATIENT WITH PEPTIC ULCER DISEASE].

Peptic ulcer disease is represented by a lesion in the mucosa of the digestive tract due to imbalance of its aggressive and protective mechanisms. The main external factors of the development of peptic ulcers are Helicobacter pylori infection and the use of non-steroidal anti inflammatory drugs (NSAIDs) and acetylsalicylic acid (ASA). Symptoms of peptic ulcer disease are a common reason for visiting the family physician. All patients with symptoms of dyspepsia under the age of 50 and without the alarm symptoms should be tested whether H. pylori is present by performing the Urea Breath Test or stool antigene testing, and infection, if found, should be treated. Endoscopic examination is obligatory in patients older than 50 years and those with alarm symptoms. "Sequential therapy" is recommended in Croatia as the first-line treatment of H. pylori infection, or triple therapy that comprises applying a proton pump inhibitor (PPI) in combination with amoxicillin and metronidazole. Four weeks after eradication therapy the control testing for H. pylori should be performed.

[NON-ALCOHOLIC FATTY LIVER DISEASE].

With the increasing prevalence of obesity and metabolic syndrome, non-alcoholic fatty liver disease (NAFLD) has become the most common liver disease in adults and children. Despite the increasing prevalence, NAFLD remains largely undiagnosed and untreated in routine medical practice. Most patients with NAFLD have no symptoms, while only a few feel discomfort in the upper right quadrant of the abdomen or increased fatigue. The diagnosis is usually set during abdominal ultrasound examination, which is done for some other reason, or by elevated liver biochemical test findings. Early recognition is very important for correct and successful treatment. In primary health care, little is known about the processes related to the identification, diagnosis and referral of patients to specialists. Differentiation between steatosis and steatohepatitis by assessing the severity of fibrosis within steatohepatitis is extremely important. Histopathologic analysis of tissue obtained by biopsy remains the gold standard in this field. For family physician, guidelines that should be combined can be of great help in treating patients with suspected NAFLD. The serious consequences of late recognition of NAFLD could be reduced by such guidelines. Recent studies have shown that NAFLD is associated with an increased prevalence and incidence of cardiovascular complications. Because of their complexity, hepatic and extrahepatic complications, heterogeneity in clinical presentation, histologic severity, prognosis and therapeutic outcome, NAFLD requires a multidisciplinary approach with the active role of family physicians in preventive care, diagnosis and treatment, especially in individuals and groups at risk. Greater attention should be focused on lifestyle modifications (reduction in body weight and physical activity) and their practical implementation. Key words: non-alcoholic fatty liver disease, liver biopsy

[CHOLESTASIS AND INFLAMMATION OF THE PANCREAS IN FAMILY MEDICINE].

Cholestasis indicates stagnation of bile, a disorder in the synthesis, secretion and/or outflow of bile. Cholestasis is classified as intrahepatic or extrahepatic. Intrahepatic cholestasis may occur as a result of hepatocellular disorders or due to obstruction of the intrahepatic bile ducts. Extrahepatic cholestasis is caused by obstruction of the bile ducts outside the liver. Cholestasis manifests as acute or chronic (>6 months). Early biochemical markers of cholestasis include increased alkaline phosphatase, γ-glutamyltransferase, and conjugated hyperbilirubinemia. Clinically, the most common presenting symptoms are jaundice, itchy skin and fatigue. A key element in diagnosis is visual presentation of the biliary system. New insights into the pathophysiological mechanisms of cholestasis at the molecular level will contribute to the development of new treatments. Pancreatitis is an inflammatory process in the tissue of the pancreas caused by prematurely activated pancreatic enzymes. The course of acute pancreatitis is in most cases mild, with minimal organ dysfunction and full recovery, while the severe form of the disease is characterized by complications and high mortality. Chronic pancreatitis is characterized by persistent inflammation of the pancreas with permanent damage to glandular tissue in the presence of fibrosis and narrowed pancreatic ducts. Treatment of pancreatitis is supportive and directed to maintaining the basic physiological functions. The role of family physicians is very important in early detection of liver diseases, which can significantly affect the success of treatment and improve the quality of life in these patients.

Comparative study on Allium schoenoprasum cultivated plant and Allium schoenoprasum tissue culture organs antioxidant status.

This study was designed to examine Allium schoenoprasum tissue culture organs antioxidant and scavenging activity and to make a comparison between Allium schoenoprasum cultivated plant and Allium schoenoprasum tissue culture organs antioxidant activity. This study reports the results on the root, stalk and leaf antioxidant enzyme activities (superoxide dismutase, catalase, guaiacol peroxidase and glutathione peroxidase), reduced glutathione quantity, flavonoids and soluble protein contents and quantities of malonyldialdehyde and ·OH radical. In Allium schoenoprasum tissue culture organs the total antioxidant capacity was determined by the FRAP method and scavenger activity by the DPPH method. The present results indicated that the crude extract of Allium schoenoprasum tissue culture exhibited antioxidant and scavenging abilities in all investigated plant parts, especially in the roots. According to our results, the tissue culture plants exhibited the highest activities in the roots in contrast to the cultivated plants where highest activities were observed in the leaves.

Systolic right ventricular adaptive changes in athletes as predictors of the maximal functional capacity: a pulsed tissue Doppler study.

AIM: The aim of this study was to extend the analysis of the systolic right ventricular (RV) adaptation to combined endurance and strength training, to assess the utility of tissue Doppler imaging in detecting the degree of these changes and to find independent RV predictors of the maximal functional capacity. METHODS: Standard Doppler and TDI were used to assess cardiac parameters at rest in 37 elite male athletes (16 wrestlers, 21 water polo players) and 20 sedentary subjects of similar age. Progressive maximal test on treadmill was used to assess VO2max. The obtained parameters were adjusted for HR, FFM, and BSA. RESULTS: Wrestlers showed higher VO2max than controls, but lesser than water polo players. RV diameter was larger in athletes. Right atrial pressure (RVE/e) was higher in water polo players than in other groups. Systolic function assessed by tricuspid annular plane systolic excursion (TAPSE) and RVs' was the highest in wrestlers. Global RV systolic parameters myocardial performance index (MPI) and preejection time/ejection time index (PET/ET) were similar. On multivariate analysis systolic parameters were independent predictors of VO2max only in wrestlers: RVs' (beta=3.18, P=0.001) and RV ET (beta=2.32, P=0.001). RVE/e` correlated with RVs' (r=-0.57, P=0.000). TAPSE correlated with RV ET (r=0.32, P=0.015) and RVs` (beta=0.28, P=0.033). CONCLUSION: Systolic function assessed by TAPSE and RVs` has more improved in less endurance athletes. RVs`and TDI ejection time predict VO2max in wrestlers, and possibly in other athletes with lesser right atrial pressure. TDI enables quantifying RV adaptation degree in athletes, but complementary to M-mode technique.

Survivin expression in odontogenic keratocysts and correlation with cytomegalovirus infection.

OBJECTIVES: The aim of this study was to investigate the expression of survivin, an inhibitor of apoptosis, in odontogenic keratocysts and to compare it to the findings in non-neoplastic jaw cysts - periapical cysts, as well as to establish a possible relationship between survivin expression and human cytomegalovirus presence within these cysts. MATERIALS AND METHODS: Samples of 10 odontogenic keratocysts (five positive and five negative for the presence of cytomegalovirus, as determined by polymerase chain reaction) and 10 periapical cysts (five positive and five negative for the cytomegalovirus presence) were analysed. The expression of survivin was assessed by immunohistochemical methods, using monoclonal antibody that selectively recognizes the cytoplasmic form of survivin. RESULTS: All 10 odontogenic keratocysts showed immunostaining for survivin, while all 10 periapical cysts were negative for its presence. There was no correlation between cytomegalovirus presence and expression of survivin within odontogenic keratocysts. CONCLUSION: Survivin may contribute to the aggressive behavior of odontogenic keratocysts, and thus support the emerging opinion of their neoplastic nature.

The utility of 99mTc-EDDA/HYNIC-TOC scintigraphy for assessment of lung lesions in patients with neuroendocrine tumors.

UNLABELLED: Our aim was to assess clinical utility of 99mTc-EDDA/HYNIC-TOC scintigraphy for evaluation of lung lesions in patients with neuroendocrine tumors (NETs). Single photon emission computed tomography (SPECT) of the thorax and whole body scintigraphy were performed in 34 patients using 99mTc-EDDA/HYNIC-TOC. Visual assessment was complemented by semiquantitative evaluation based on tumor to non-tumor (T/NT) ratio. Clinical, laboratory, and histological findings served as the standard for comparison. Enhanced tracer uptake was observed on both SPECT and whole body scintigraphy in 29 of 34 patients (88% sensitivity). T/NT ratios were significantly higher on SPECT than whole body images (2.96+/-1.07 vs.1.70+/-0.43, p KEYWORDS: 99mTc-EDDA/Hynic-TOC, lung involvement of NETs, T/NT ratio.

Comparison between polyphenol profile and bioactive response in blackthorn (Prunus spinosa L.) genotypes from north Serbia-from raw data to PCA analysis.

The aim of this work was to investigate and compare the phenolic profile of 15 wild growing blackthorn (Prunus spinosa L.) genotypes from the slopes of Fruska Gora mountain in north Serbia. Their effect in inhibiting i) α-amylase and α-glucosidase activities and ii) colorectal cancer cell line (HT29) growth was also studied. Blackthorn fruit extracts exhibited high phenolic content being enrich in anthocyanins. Principal component analysis was used to correlate the bioactive response with phenolic composition. It was found that derivatives quercetin and anthocyanin peonidin are the major contributors of the inhibition of carbohydrates hydrolyzing enzymes as well as with the antiproliferative effect of blackthorn. Among all samples, the genotype from Beska locality showed the higher capacity in inhibiting alpha-amylase, alpha-glucosidase and HT29 cell growth. Because of high anthocyanin content and higher bioactive response, these genotypes could be recommended for the further cultivation and investigation.

Antioxidant capacity of Melampyrum barbatum--weed and medicinal plant.

This study was designed to examine antioxidant and free radical scavenging activities of red and yellow forms of Melampyrum barbatum L. In this study, we report the results concerning flower and leaf antioxidant enzyme activities (superoxide dismutase, catalase, guaiacol peroxidase and glutathione peroxidase), reduced glutathione quantity, flavonoids, photosynthetic pigments and soluble protein contents and quantities of malonyldialdehyde, (*)OH and O(2) (*-) radicals; total antioxidant capacity was determined by FRAP (Ferric Reducing Antioxidant Power) method and scavenger activity by DPPH (1,1-diphenyl-2-pycril-hydrasil radical) method. Lipofuscin 'plant age pigments' were also determined. According to our results, flowers of the red form of M. barbatum exhibited the highest antioxidant capacity.

Free radical-decellularized tissue promotes enhanced antioxidant and anti-inflammatory macrophage response.

Oxidative stress leads to the progression of many diseases including chronic wounds, atherosclerosis, stroke and cancer. The modification of biomolecules with reactive nitrogen or oxygen species has been shown to trigger oxidative stress pathways that are beneficial for healing. Extracellular matrix scaffolds have been used successfully in reconstructive applications due to the beneficial host response they induce. To tailor extracellular matrix scaffolds to enhance antioxidant response, ECM were prepared using reactive nitrogen or oxygen species. These scaffolds were shown to be effectively decellularized and possess oxidative or nitroxidative protein modifications. Macrophage responses in vitro and in an in vivo muscle injury model were shown to have enhanced antioxidant phenotypes without impairment of long-term remodeling. These observations suggest that ECM decellularized with reactive oxygen or nitrogen species could provide better outcomes for the treatment of ischemic diseases.

Antioxidant and free-radical scavenging activities of Allium roseum and Allium subhirsutum.

This study was designed to examine the antioxidant and free-radical scavenging activities of two Allium species, Allium roseum and Allium subhirsutum. This study reports the results concerning bulb, stalk and leaf antioxidant enzyme activities (superoxide dismutase, catalase, guaiacol peroxidase and glutathione peroxidase), quantities of malonyldialdehyde, (*)OH and O(2) (*-) radicals and total antioxidant capacity determined by the FRAP method. Scavenging activities were determined by ESR method. The total antioxidant capacity was the highest in the leaves of Allium roseum. Among the investigated organs the leaves exhibited the highest antioxidant activities in all investigated Allium species. The highest ESR (84.61%) scavenger activity was observed in the leaves of Allium roseum.

Antioxidant and scavenger activities of Allium ursinum.

The antioxidative properties of bulb, leaf and stalk of Allium ursinum were investigated. Activities of antioxidant enzymes (superoxide dismutase, catalase, peroxidase, glutathione peroxidase), quantities of malonyldialdehyde, superoxide and hydroxyl radicals and reduced glutathione and also the contents of total flavonoids, chlorophylls a and b and carotenoids were determined. The extracts from all plant organs exhibited antioxidant activity, the highest having been observed in the leaves. Furthermore, ESR signal of PBN-OH radical adducts in the presence of leaves phosphate buffer (pH 7) extract was reduced for 87.61%.

Psycho-social model of depression within the elderly population in the institutional and non-institutional life conditions.

The aged people are often in need of other people's help available at their home or in institutions for long term care. On the other hand depression is one of the most prevalent psychiatric disorders and a common cause of disability in the elderly. The study included N=100 participants. Subjects are individuals in the age of 65 or more, 39 men and 61 women. Fifty subjects have been accommodated in the Elderly Centres (homes for retirees), and fifty have been using gerontology centre services at their homes. Depression, functional capability, psychosomatic disorders and subjective estimate of health have been investigated. It has occurred that depression in general was more present among the subjects who lived in the institutional life conditions, namely that depression major (major, regarding severity of the disease) is more frequently present in this group. Depression minor is more frequent among those subjects who live in the non-institutional conditions. Psychosomatic disorders, decreased functional capability and lack of contacts outside homes are in larger correlation to depression among subjects living in the non-institutional life conditions. Depression and subjective health estimate are highly correlated in both groups. Depression is more widely present among women and subjects with higher education levels.

Secular trend in growth and nutritional status in a sample of girls aged 7-9 years from Serbia.

Secular trend changes over time and therefore is a marker of the public health of the populations. The aim of the study is to compare the anthropometric data of 7-9 year-old girls obtained in recent investigations in Serbia with data from 1980/81, by using Z-score and percentile classification systems, according to the Center for Disease Control and Prevention - CDC anthropometric criteria. In 1980 and 1981 we used data for 800 schoolgirls aged 6.50-9.49 years. The data for the period 2011-2016 were obtained from recently conducted field investigations in certain parts of the Vojvodina region and from the East and Central Serbia. In total, 1313 girls were included in both investigations. Mean Z-score values for height-for-age, weight-for-age and BMI-for-age in the present sample were compared with the corresponding values for the period 1980/81, and also compared with the reference. The results show that nutritional status Z-scores indicate similar mean values in both investigations. The secular trend of height has slowed down, but the percentage of girls with normal weight is considerably lower in the second investigation. The results indicate that nutritional problems are more common among present generation of girls. Both the problems of underweight and overweight in girls aged 7-9 have to be addressed.

Bcl-2 expression in oral squamous cell carcinoma.

Apoptosis is a genetically regulated process involved in tissue size regulation, morphogenesis, and elimination of genetically damaged cells. A pallet of genes is involved in the control of apoptosis, such as bcl-2 family whose oncogenic potential has been demonstrated in oral tumorigenesis. Different members of bcl-2 family may promote or inhibit apoptosis by synthesizing anti- and proapoptotic proteins. One of antiapoptotic proteins, bcl-2, with a crucial role in apoptosis regulation was the object of our study. By means of immunohistochemistry we estimated the level of overexpression of bcl-2 proteins in a series of the 26 formalin fixed, paraffin-embedded samples of oral squamous cell carcinoma (OSCC). Analyzed tumors originated from different sites of oral cavity; 7/26 belonged to stage II, 14/26 to stage III, and 5/26 to stage IV. Immunoreactivity was scored according to the percentage and intensity of positive cytoplasmic bcl-2 staining. All tumors had low percentage of positively stained bcl-2 cells, with mean values for lower/higher intensity of 8.3 +/- 2.5/34.4 +/- 7, 7.5 +/- 1.1/31.9 +/- 4.3, and 8.4 +/- 5.8/31.5 +/- 5.8 within stages II, III, and IV, respectively. Low level of bcl-2 expression in our sample seems to be associated with higher survival rate: 77% for the 5-year follow-up period. Comparing clinicopathologic and risk factors data within each and between three groups of analyzed tumors (lip-tongue P = 0.58, tongue-floor of the mouth, P = 0.21, lip-floor of the mouth, P = 0.50) there was no significant difference. However, our results suggest that the level of bcl-2 expression could be a valuable predictor of tumor behavior and disease outcome.

[Long-term clinical benefits of using active stents in diabetic patients: two year follow-up of a consecutive series of 122 patients]. / Bénéfice clinique à long terme de l'utilisation des stents actifs chez les patients diabétiques: suivi à deux ans d'une série consécutive de 122 patients.

The objective of our study is to evaluate the long-term results of coronary angioplasty using active stents in a population of diabetic patients. This is a single-centre study on a consecutive series of 122 diabetic patients (40% of them insulin dependent) who between January 2003 and June 2004 underwent angioplasty with implantation of an active stent (sirolimus Cypher(R) or paclitaxel Taxus(R)) for one or more de novo coronary lesions. The mean age was 66 +/- 10 years and a total of 171 coronary segments were treated. The lesions treated were complex (type B2 + C) in 69% of the cases, with a mean stent length of 21 +/- 15 mm and a mean stent diameter of 2.7 +/- 0.3 mm. Follow-up at two years for 119 patients (3 lost to follow-up) revealed a mortality rate of 4.2%, and a myocardial infarction rate of 7.5%. The rates for revascularisation of the target lesion and the target vessel were 11.4% and 17.8% respectively, with a rate of major cardiac events of 22.5%. During this period, 25.2% of the patients underwent revascularisation of at least one vessel. This study confirms the benefits of using active stents for revascularisation of the target lesion in diabetic patients. However, it serves as a reminder that the progression of coronary atheroma is global, and that the prognosis for these patients depends essentially upon managing risk factors, and particularly on controlling their diabetes.