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.

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.

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.

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.

The effects of specific exercise program on anthropometric characteristics and motor abilities of preschool children / Efectos del programa de ejercicios específicos sobre las características antropométricas y las habilidades motrices de los niños de edad preescolar

In order to determine the effects of specific exercise program on anthropometric characteristics and motor abilities of preschool children, a battery of eight anthropometric measures and 7 motor tests were applied. The current study was conducted on a sample of 60 preschoolers (31 in the experimental and 29 in the control group), aged 4 to 5. They were the respondents in the project implemented by the Faculty of Sport and Physical Education in Novi Sad. The obtained data was analyzed by multivariate analysis of covariance (MANCOVA). It can be concluded that the application of experimental and control models of exercise resulted in substantial improvement of certain motor skills in the experimental group compared to the control group. In regard to the anthropometry there was no statistically significant difference between the groups. In fact, the practical value of this work is to analyze the model of kinetic activities, which as such can be applied to all clubs and schools that work with children of preschool and early school age.

Con el fin de determinar los efectos de un programa de ejercicios específicos sobre las características antropométricas y las habilidades motrices de los niños en edad preescolar, se aplicó una batería de 8 medidas antropométricas y 7 pruebas motrices. El presente estudio se realizó en una muestra de 60 niños en edad preescolar (31 en el grupo experimental y 29 en el grupo de control), de 4 a 5 años. Los niños en este estudio forman parte del proyecto implementado por la Facultad de Deporte y Educación Física de Novi Sad. Los datos obtenidos se analizaron mediante análisis multivariante de covarianza (MANCOVA). Se puede concluir que la aplicación de modelos experimentales y de control de ejercicios resultó en una mejora sustancial de ciertas habilidades motoras en el grupo experimental en comparación con el grupo control. En cuanto a la antropometría, no hubo diferencias estadísticamente significativas entre los grupos. De hecho, el valor práctico de este trabajo es analizar el modelo de actividades cinéticas, que como tal puede aplicarse a todos los clubes y escuelas que trabajan con niños en edad preescolar y escolar.

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.

Structural Characterisation Reveals Mechanism of IL-13-Neutralising Monoclonal Antibody Tralokinumab as Inhibition of Binding to IL-13Rα1 and IL-13Rα2.

Interleukin (IL)-13 is a pleiotropic T helper type 2 cytokine frequently associated with asthma and atopic dermatitis. IL-13-mediated signalling is initiated by binding to IL-13Rα1, which then recruits IL-4Rα to form a heterodimeric receptor complex. IL-13 also binds to IL-13Rα2, considered as either a decoy or a key mediator of fibrosis. IL-13-neutralising antibodies act by preventing IL-13 binding to IL-13Rα1, IL-4Rα and/or IL-13Rα2. Tralokinumab (CAT-354) is an IL-13-neutralising human IgG4 monoclonal antibody that has shown clinical benefit in patients with asthma. To decipher how tralokinumab inhibits the effects of IL-13, we determined the structure of tralokinumab Fab in complex with human IL-13 to 2 Å resolution. The structure analysis reveals that tralokinumab prevents IL-13 from binding to both IL-13Rα1 and IL-13Rα2. This is supported by biochemical ligand-receptor interaction assay data. The tralokinumab epitope is mainly composed of residues in helices D and A of IL-13. It is mostly light chain complementarity-determining regions that are driving paratope interactions; the variable light complementarity-determining region 2 plays a key role by providing residue contacts for a network of hydrogen bonds and a salt bridge in the core of binding. The key residues within the paratope contributing to binding were identified as Asp50, Asp51, Ser30 and Lys31. This study demonstrates that tralokinumab prevents the IL-13 pharmacodynamic effect by binding to IL-13 helices A and D, thus preventing IL-13 from interacting with IL-13Rα1 and IL-13Rα2.

The Parasol Protocol for computational mutagenesis.

To aid in the discovery and development of peptides and proteins as therapeutic agents, a virtual screen can be used to predict trends and direct workflow. We have developed the Parasol Protocol, a dynamic method implemented using the AMBER MD package, for computational site-directed mutagenesis. This tool can mutate between any pair of amino acids in a computationally expedient, automated manner. To demonstrate the potential of this methodology, we have employed the protocol to investigate a test case involving stapled peptides, and have demonstrated good agreement with experiment.

Water stress induces changes in polyphenol profile and antioxidant capacity in poplar plants (Populus spp.).

This paper is aimed to characterize young poplar plants under the influence of water stress provoked by polyethileneglycol 6000 (PEG 6000). Three polar genotypes (M1, B229, and PE19/66) were grown in hydroponics and subjected to 100 and 200 mOsm PEG 6000 during six days. Polyphenol characterization, two enzymatic markers and antioxidant capacity in leaves and roots were investigated in stressed plants. Total phenol content, ferric reducing antioxidant capacity (FRAP) and DPPH antiradical power (DPPH ARP) were determined for estimating total antioxidant capacity. Polyphenol oxidase (PPO) and phenylalanine ammonia lyase (PAL) were determined as enzymatic markers. Polyphenol characterization of poplar samples was performed by HPLC-PDA analysis. All results were subjected to correlation analysis and principal component analysis (PCA). Inspite of the decrease of total phenol content in investigated genotypes, as well as total antioxidant capacity, some of polyphenols were affected by stress like flavonoids chrysin, myricetine, kaempferol and isoferulic acid in roots of B229 genotype (Populus deltoides). Genotype B229 also showed the increase of antioxidant capacity and PAL activity in root and leaves under stress what could be the indicator of the adaptability of poplar plants to water stress. Significant positive correlations were obtained between PAL, antioxidant capacity as well as phenolic acids among themselves. Chemometric evaluation showed close interdependence between flavonoids, FRAP, DPPH antiradical power and both investigated enzymes of polyphenol metabolism, PAL and PPO.

Superior lung capacity in swimmers: Some questions, more answers!

BACKGROUND: Physical activity has a positive effect on the function of the whole human body system. The influence of physical activity on the development of the respiratory system is still a matter for debate. Swimming is considered the sport with the most profound effect on the lungs. AIM: The first aim was to determine pulmonary function and to correlate it with anthropometric features of sportsmen, represented by land- and the water-based elite athletes comparing with their sedentary counterparts; the second aim was to examine whether the training factors (frequency and amount) influence pulmonary function in swimmers, when controlled for anthropometric features. METHODS: Thirty-eight elite male swimmers were matched for age and sex with two hundred and seventy-one elite football players and one hundred controls who were not involved in any routine exercise. Lung volumes were recorded by Pulmonary Function test and analyzed statistically. RESULTS AND CONCLUSION: Swimmers had statistically higher values of VC, FVC, FEV1 and FEV1/FVC when compared to both the football players and the controls, as the latter two showed no in-between differences. There was significant positive correlation between age, body weight and body height and each of the above named pulmonary parameters, when presented separately for swimmers, football players and the control group. When controlled for the anthropometric features, larger lung volumes in swimmers were not influenced by training period, age at the beginning of training and weekly extent of personal training. Further comprehensive longitudinal studies are needed to confirm these observations.

The amplification of c-erb-B2 in cancer-free surgical margins is a predictor of poor outcome in oral squamous cell carcinoma.

The tumour subtype, TNM classification, and histopathological data are sometimes not sufficient for understanding and assessing the behaviour of oral cancers. In an attempt to find additional markers of tumour biology and behaviour, this study sought to determine the incidence and consequently the relevance of c-erb-B2, c-Myc, and H-ras gene alterations in tumour-free margins of oral squamous cell carcinoma (OSCC). Fifty samples of OSCC were analyzed for c-erb-B2 and c-Myc amplification by real-time polymerase chain reaction and for H-ras point mutations by sequencing. A relatively high incidence of genetic lesions was detected: 22% of cases had c-erb-B2 and 30% had c-Myc amplification, whilst only 12% harboured H-ras mutations. Kaplan-Meier analysis and the log-rank test showed statistically significant differences in 5-year survival rates and relapse between patients with tumour margins positive for c-erb-B2 amplification and those with margins that were negative (P=0.002). H-ras and c-Myc alterations could not be associated with tumour behaviour. Molecular analysis of margins, targeting cancer genes, could identify additional, independent predictors of risk and outcome in OSCC.

[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.

[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.

[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.

Evaluation of central corneal thickness in patients with ocular hypertension and primary open-angle glaucoma.

AIM: To determine correlation between central corneal thickness (CCT) and applanation intraocular pressure (IOP) in normal patients with primary open angle glaucoma and patients with ocular hypertension. METHODS: Two-year retrospective study designed to determine correlation between CCT and IOP. Records of 121 patients were included in the study. Inclusion criteria were: IOP higher than 22 mm Hg for a group with ocular hypertension but without functional, morphological damage and progression, diagnosis of glaucoma for a group with open-angle glaucoma and normal ophthalmological results for a control group. Patients who were mono-oculus, patients with secondary glaucoma and who had surgical treatment were excluded. The patients were selectively grouped according to types of glaucoma, by gender and age. The IOP values were measured by Goldman's applanation tonometer, CCT values were determined using ultrasonic pachymeter, in the period from January 2011 to December 2012. RESULTS: Of total 121 subjects, 51 had primary open angle glaucoma (POAG), 40 had ocular hypertension (OHT) and 30 had normal ophthalmological findings (control group). The CCT values in OHT group 529.37+/-25.18 µm were greater than of both POAG, 501.02+/-14.04 µm and control group, 497.37+/-14.90 µm. The IOP values in OHT group were 20.25+/- 3.62 mm Hg, and in POAG group were 18.25+/-2.70 mm Hg, while in the control group they were 13.53+/- 3.62 mm Hg. CONCLUSION: Determination of CCT in patients with glaucoma is crucial, considering its impact on IOP values, which represents a parameter in glaucoma diagnosis, as well as monitoring further progress of ocular hypertension and patients diagnosed with glaucoma.

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.

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.