Nitric oxide synthase expression and activity in cell models relevant to human pathophysiology

Složene interakcije mitohondrijske i nuklearne DNA onemogućuju otkrivanje pravog uzroka nasljednih mitohondrijskih bolesti kao i pružanje učinkovite terapije. Leberova nasljedna optička neuropatija (LHON), jedna od najučestalijih nasljednih optičkih neuropatija, je posljedica točkastih mutacija mitohondrijske DNA koje dovode do smanjenja aktivnosti kompleksa I u respiratornom lancu. Tipična obilježja LHON-a, sa biokemijskog stajališta, su smanjenje proizvodnje energije na staničnoj razini i povećana proizvodnja reaktivnih kisikovih (ROS) i dušikovih (RONS) spojeva no, to ne objašnjava odgođenu pojavu simptoma bolesti i učinak samo na određene stanice. Naime, iako je mutacija prisutna u cijelom tijelu, štetan učinak iskazuje samo na ganglijskim stanicama mrežnice. Obzirom da je LHON karakteriziran deficitom oksidativne fosforilacije, a NO· je poznati negativni modulator mitohondrijskog respiratornog lanca istražili smo njegovu ulogu u razvoju LHON-a. Rezultati u ovom radu pokazuju da dugotrajna izloženost dušikovom oksidu ima veći utjecaj na smrtnost stanica LHON-a uspoređujući je sa smrtnosti kontrolnih stanica i pretpostavljaju da se ROS i RONS stvaraju u većim količinama u ovom patološkom stanju i vode ka staničnoj smrti. Vođeni tom mišlju, provjerili smo i količinu iNOS-a i eNOS-a na razini mRNA. Obzirom da rezultati nisu statistički značajni, pretpostavili smo da postoji razlika u ekspresiji jednog ili više NOS enzima kod pacijenata oboljelih od LHON-a ali ovo pitanje ostaje otvoreno za daljnja istraživanja. Buduća perspektiva je potpuno objašnjenje uloge NO· u specifičnim mitohondrijskim bolestima i rješavanje zagonetne ekspresije i aktivnosti samog NOS-a. Možda upravo metabolizam NO· može biti važan u patofiziologiji LHON-a te njegovo razumijevanje može pridonijeti razvoju efikasnijih terapija i preventivnih mjera.Due to complex interactions of mitochondrial and nuclear DNA, the real cause of inherited mitochondrial diseases is still unclear and patients currently lack effective treatments. Leber’s hereditary optic neuropathy (LHON), one of the most common inherited optic neuropathies, is caused by point mutations in mtDNA that result in a defect of respiratory chain complex I activity. From the biochemical point of view the bioenergetic failure and the enhanced reactive oxygen species (ROS) production are typical hallmarks of LHON disease, but still they do not explain the late onset and the incomplete penetrance. Moreover it is still unknown why, although the mutation is present overall the body only the RGCs are affected. Since LHON is characterized by a deficit in the oxidative phosphorylation chain and NO· is an established mitochondrial respiratory chain negative modulator, we checked if it might have a role in the LHON development and progression. The results presented in this thesis prove that long–term exposure to NO· has a larger impact on LHON cells viability compared to controls and suggest that higher levels of reactive oxygen and nitrogen species (ONOO-) are produced in this pathological state, leading to cell death. In addition, careful though still statistically insufficient evaluations of the levels of iNOS and eNOS mRNA expression seems to suggest that there might be a difference in the expression of one or more NOSs in LHON patients: this question however remains open. Future perspectives are, indeed, to provide a complete picture about involvement of NO· in this specific mitochondrial disease and to solve the conundrum of the NOSs expression and activity. The feeling is that the NO· metabolism can be relevant to LHON pathophysiology, its comprehension opening to the development of more efficient therapeutic and preventive interventions

Serum visfatin concentration in eutrophic and overweight/obese male children in early childhood

Background and Purpose: Childhood overweight/obesity is considered a global epidemic, which began earlier in pediatric patients and presents a major health risk in adulthood. It is well known that adipose tissue is an active endocrine inflammatory organ in obese adults, but its neurohormone activity in the childhood is not yet clarified. Visfatin is one of adipokines with insulin-mimetic and proinflammatory-atherosclerotic effect, whose role in the child’s age is still unknown as well as its physiological concentrations in serum of prepubertal children. The aim of this study was to determine visfatin serum concentration at the early age in eutrophic and overweight/obese male children and its association with arterial blood pressure. Materials and Methods: Healthy boys, 2-14 years old, hospitalized for elective inguinal hernia surgery has been included in the study (N=31) and were divided into two groups according to percentile curve: a) overweight/obese (O/O:above the 85th percentile) and b) eutrophic (E:5th-85th percentile). Anthropometric and biochemical measurements and specific serum levels of visfatin by enzyme immunoassay were determined. Results and Conclusion: Both groups of examinees, eutrophic and overweight/obese, had a normal metabolic profile band on percentile ranks (fasting glucose values and all fractions of lipid profile) and values of blood pressure. However overweight/obese boys had significant higher systolic blood pressure than eutrophic boys, p=0,028. Serum visfatin were 6.90±3.97 ng/ml in eutrophic boys, while in overweight/obese were 7.82±3.75 ng/ml, p=0,57. There is a tendency of visfatin serum concentration to increase with increase of body weight and growing. This suggests possible role of visfatin in future metabolic and cardiovascular processes related to increase in body mass

TESTING OF NATURAL STONE - SLIP RESISTANCE USING A PENDULUM

Ovaj rad prezentira ispitivanje otpornosti kamena na klizanje. Ispitivanje se provodi prema normi BAS EN 14231:2009, koja utvrđuje metodu za određivanje vrijednosti otpornosti na klizanje, površine izloženog lica elemenata od prirodnog kamena, namijenjenih za oblaganje podova u građevinarstvu. Cilj rada je prikazati opremu, pomoćna sredstva, uzorke i provedbu ispitivanja. Ispitivanja se provode na uzorcima kamena s različitim završnim obradama. Ispitne površine su: rezana, polirana i površina dobivena paljenjem. Nakon provedenih ispitivanja autori analiziraju i vrednuju dobivene rezultate. U sklopu 4. međunarodnog simpozija Hercegovina zemlja kamena prvi put su predstavljeni rezultati ovog stručno-znanstvenog istraživanja.This paper presents an examination of natural stone to slip resistance. The test is carried out according to the BAS EN 14231:2009 standard. This standard specifies a test method for determining the slip resistance value of the surface of the exposed face on natural stone elements intended to be used for flooring in buildings. The aim of the paper is to present the equipment, auxiliary means, samples and the test procedure. Tests are carried out on stone samples with different finishes. The test surfaces are: cut, polished and flamed. After the tests, the authors analyze and evaluate the obtained results. The results of this scientific research were presented for the first time as part of the 4th international symposium Herzegovina Land of Stone

Urine neutrophil gelatinase-associated lipocalin concentration in healthy newborns during the first three postnatal days

Introduction: Urine neutrophil gelatinase-associated lipocalin (uNGAL) is a biochemical marker significant for early prediction of acute kidney injury in adults. However, it has not been examined sufficiently among the infant population, particularly newborns in terms of reference values. The aim of our study was to determine the concentration of uNGAL in healthy term newborns and to determine if there was a difference in uNGAL concentration according to gender, postnatal age and birth weight. Materials and methods: Our study involved 81 healthy term newborns birth (≥ 37 weeks, Apgar score ≥ 8 in the first minute after birth, CRP < 5 mg/L). Urine NGAL was measured using chemiluminescent microparticle immunoassay (CMIA) within 72 hours after birth, on Architect plus ci8200 analyser (Abbott, Chicago, USA). Data were analysed using Statistica software. Results: The median concentration of uNGAL in the whole study group of healthy term newborns was 27.1 ng/mL (16.5-56.0 ng/mL) (newborn girls, 27.1 ng/mL (15.8-47.9 ng/mL); newborn boys, 27.9 ng/mL (16.5-61.0 ng/mL), P = 0.941). Median uNGAL concentration according to postnatal age expressed in days was 28.2 ng/mL (11.7-57.2 ng/mL) 1st day, 28.9 ng/mL (16.5-64.2 ng/mL ) 2nd day and 23.9 ng/mL (20.2-46.6) 3rd day, P = 0.863. Regarding birth weight for newborns < 3500 g, median concentration was 25.0 ng/mL (16.5-45.4 ng/mL ) and for weight ≥ 3500 g 30.6 ng/mL (16.5-64.2 ng/mL), P = 0.455. Conclusions: There were no significant difference in uNGAL concentration in relation to gender, postnatal age and birth weight

Hyperleptinemia – Non-Haemodynamic Risk Factor for the Left Ventricular Hypertrophy Development in Hypertensive Overweight Females

Obesity is directly and strongly associated with hypertension and left ventricular hypertrophy (LVH). Development of LVH is multifactorial, caused both by haemodynamic and non-haemodynamic factors. Hypertension is the main haemodynamic factor. Humoral mechanisms, as a non-haemodynamic factor for LVH development, have not been completely explained. The aim of this study is to determine whether hyperleptinemia can be one of humoral – non-haemodynamic factor inducing LVH together with haemodynamic factors in overweight females. The study was done on thirty six adult, overweight female patients, body mass index in range 25–30 kg/m2. Patients are nondiabetic with regular renal function. Twenty one female patients were hypertensive with left ventricular hypertrophy. Control group included fifteen hypertensive female patients without left ventricular hypertrophy. In all patients was determined glucose profile and creatinine clearance, cholesterol, triglycerides, LDL, HDL.Weight, high, circumference of the waist and hips was taken. Cardiovascular determination was done applying two-dimensional ultrasound. Serum leptin level was measured using radioimmunoassay method (RIA). Results showed that serum leptin level was significantly higher in hypertensive, overweight females with LVH. This suggests that non-haemodynamic factors, such as hyperleptinemia, participate in left ventricular hypertrophy development together with haemodynamic factors in adult hypertonic, overweight females

Evidence for detrimental cross interactions between reactive oxygen and nitrogen species in Leber's hereditary optic neuropathy cells

Here we have collected evidence suggesting that chronic changes in the NO homeostasis and the rise of reactive oxygen species bioavailability can contribute to cell dysfunction in Leber’s hereditary optic neuropathy (LHON) patients.We report that peripheral blood mononuclear cells (PBMCs), derived froma female LHON patient with bilateral reduced vision and carrying the pathogenic mutation 11778/ND4, display increased levels of reactive oxygen species (ROS) and reactive nitrogen species (RNS), as revealed by flow cytometry, fluorometric measurements of nitrite/nitrate, and 3-nitrotyrosine immunodetection. Moreover, viability assays with the tetrazolium dye MTT showed that lymphoblasts from the same patient are more sensitive to prolonged NO exposure, leading to cell death. Taken together these findings suggest that oxidative and nitrosative stress cooperatively play an important role in driving LHON pathology when excess NO remains available over time in the cell environment

Radioactivity Measurements of Ceramic Tiles Produced in Serbia

This paper presents the results of gamma spectrometry measurements of natural radionuclides ( 226Ra, 232Th, and 40K) in some floor and wall ceramic tiles produced in Serbia and used in homes and workplaces. The level of radioactivity of some ceramic tiles produced in Serbia by two major manufacturers–Zorka Keramika and Toza Markovic was examined. The measured mean value of the activity concentration of 226Ra, 232Th, and 40K exceeds the average values in the world for building materials with values of 67.2±6.9 Bq kg-1 for 226Ra, 57.4±4.7 Bq kg-1 for 232Th and 808±48 Bq kg-1 for 40K. Based on these calculated values, the representative level index gamma index, associated with gamma radiation, whose average value is 0.78±0.06, and annual effective dose, whose average value is 0.117±0.009 mSv y-1 for home was obtained. Estimated values fulfill all the recommendations of the European Union for building materials, thus analyzed materials are considered not to be a health hazard for the public.BPU11 : 11th International Conference of the Balkan Physical Union : Proceedings book; Aug 11 - Sep 1, 2022S01-NPNE Nuclear Physics and Nuclear Energ

Low Adiponectin Serum Level – Reduced Protective Effect on the Left Ventricular Wall Thickness

Adiponectin, secreted by fat tissue, is down – regulated in obesity and may be involved in obesity-related disorders. It has anti-inflammatory, antiatherosclerotic and antidiabetic effect. Obesity is a strong predictor for hypertension and cardiovascular diseases. Recent studies showed that adiponectin level has important role in metabolic disorders, arterial hypertension and ischemic heart disease but its effect on left ventricular hypertrophy (LVH) has not been fully clarified. The aim of this research is to determine whether the protective effect of adiponectin against development of left ventricular hypertrophy is decreased in hypertensive overweight patients. The study included 61 adult, overweight hypertensive patients, with body mass index in range 25–30 kg/m2. Patients had regular morning glucose serum values and regular creatinine level. They were divided into four groups, according to sex and the presence of LVH. There were 16 female and 15 male hypertensive patients with LVH and 15 female and 15 male hypertensive patients without LVH, who were a control group. Glucose profile, lipidogram, creatinine clearance and anthropometric measures were determined in all patients. Cardiovascular measurements were taken applying two-dimensional ultrasound. Adiponectin serum level was measured using enzyme immunoassay (ELISA). Results showed that adiponectin serum level was significantly lower in hypertensive, overweight females and males with LVH than in the control groups without LVH. Adiponectin serum level did not correlate significant with intraventricular or with posterior wall thickness of left ventricle. Hypoadiponectinemia presents part of neurohumoral, non-haemodynamic system who contributes to obesity-related hypertension and left ventricular hypertrophy development. Low adiponectin level together with others adipokines, cytokines and chemokines secreted by fat tissue could contribute to pathophysiologic changes of the myocardium via unknown molecular mechanisms yet

Nonaqueous Polyethylene Glycol as a Safer Alternative to Ethanolic Propolis Extracts with Comparable Antioxidant and Antimicrobial Activity

We compared the chemical composition, antioxidant and antimicrobial activity of two propolis extracts: one obtained with nonaqueous polyethylene glycol, PEG 400 (PgEP), and the other obtained with ethanol (EEP). We analyzed the total phenolic content (TPC) and the concentrations of ten markers of propolis antioxidant activity with HPLC-UV: caffeic acid, p-coumaric acid, trans-ferulic acid, trans-cinnamic acid, kaempferol, apigenin, pinocembrin, chrysin, CAPE, and galangin. Antioxidant activity was tested using DPPH and FRAP assay, and antimicrobial activity was assessed through minimum inhibitory concentrations (MICs) and minimum biofilm eradication concentration (MBEC) determination. Maceration gave the yield of propolis of 25.2 ± 0.08% in EEP, and 21.5 ± 0.24% in PgEP. All ten markers of antioxidant activity were found in both extracts, with all marker concentrations, except kaempferol, higher in EEP. There was no significant difference between the TPC and antioxidant activity of the PgEP and the EEP extract ; TPC of PgEP was 16.78 ± 0.23 mg/mL, while EEP had TPC of 15.92 ± 0.78 mg/mL. Both extracts had antimicrobial activity against most investigated pathogens and Staphylococcus aureus, Acinetobacter baumannii, and Escherichia coli biofilms. EEP was more effective against all tested susceptible pathogens, except E. coli, possibly due to higher content of kaempferol in PgEP relative to other polyphenols. Nonaqueous PEG 400 could be used for propolis extraction. It gives extracts with comparable concentrations of antioxidants and has a good antioxidant and antimicrobial activity. It is a safe excipient, convenient for pediatric and veterinary formulations

Plasma leptin in obesity related hypertension

Background and Purpose: It is well known that obesity is related to hypertension through several mechanisms, such as sympathetic overactivity and excess renal sodium reabsorption. Obesity and hypertensionmay also be linked by leptin, a peptide that is elevated in obese individuals. Leptin is an adipocyte derived hormone that acts in hypothalamus to regulate appetite, energy expenditure and sympathetic nervous system outflow, as well as in peripheral tissues, such as blood vessels and kidneys. The aim of our research was to determine plasma leptin in hypertensive and normotensive obese patients with the same body mass index (BMI) and show a possible difference between leptin levels in these two groups of patients. Materials and Methods: The research was carried out on 21 hypertensive and 19 normotensive patients (20 men and 20 women) with BMI in range 30–35 kg/m2, normal values of fasting plasma glucose, urea and creatinine. Leptin was determined using Elisa method. Results and Conclusion: The results showed that men had significantly higher waist to hip ratio (W/H). Plasma leptin was significantly higher in hypertensive women as well as in hypertensive men (p=0.03) in relation to normotensive patients. There was also significant difference in serum craetinine and creatinine clearance between normotensive and hypertensive women, although creatinine was within normal range. There was a significant difference in serum tryglicerides between men and women in general, where men had higher values (p=0.016). This confirmed that leptin correlates with hypertension in both men and women