Assessment of Deoxyribonuclease Activity in Biological Samples by a Fluorescence Detection-Based Method

Objective: To develop a speedy, sensitive, low-cost fluorescence detection assay for assessing deoxyribonuclease activity in biological samples. Methods: Deoxyribonuclease activity was measured in serum samples of 15 patients with lung cancer and 15 healthy individuals. The assay is based on hydrolysis of fluorescently labeled DNA into fragments that are subsequently measured by capillary electrophoresis. Results: Mean (SD) reduction in signal intensity compared with the negative control reaction was 0.04 (0.09) for the group of patients with lung cancer and 0.01 (0.07) for the control group. The difference in signal intensity between the 2 groups was not statistically significant. Conclusion: The main advantages of the fluorescence detection assay are its high sensitivity, the short time required for it to obtain results, and its low cost. The method provides a useful alternative to the conventional enzyme-linked immunosorbent assay (ELISA) and can potentially be applied to a variety of studies, especially when assessment of deoxyribonuclease activity in large sample series is required

Predictive biomarkers for response to neoadjuvant chemoradiotherapy in colorectal cancer

Different types of biomarkers can be analyzed at various levels and by different techniques in order to assess the response to neoadjuvant chemoradiotherapy (nCRT) in colorectal cancer (CRC). The clinical practice currently relies mostly on imaging markers, carcinoembryonic antigen (CEA), and tumor histopathology. Molecules that have predictive potential in CRC patients subjected to nCRT are those involved in cellular pathways that metabolize specific chemotherapeutics and protect the cell from radiation, but none of them have been approved in clinical practice. Candidate molecules investigated so far can be sorted in the following groups based on their physiological functions: 5-FU metabolism pathways, cell cycle and DNA repair, and oxidative stress and inflammation. However, differences in schedules and doses of chemoradiotherapy regimens, variations in intervals between nCRT and surgery, and non-standardized tumor response evalution make comparisons of results among studies and drawing conclusions extremely difficult

Assessment of Deoxyribonuclease Activity in Serum Samples of Patients With Systemic Lupus Erythematosus: Fluorescence-Based Method Versus ELISA

BackgroundWe report the improvement of previously described method for determining deoxyribonuclease (DNase) activity in serum samples that uses a fluorescently labeled DNA fragment as a substrate MethodsActivity of serum DNase was analyzed in 31 patients with systemic lupus erythematosus (SLE) and 13 healthy individuals by fluoresence-based method and ELISA test ResultsWe found a mean decrease in DNase activity between cases and controls of 12.46% measured by the fluoresence-based method and of 12.21% measured by ELISA method. High level of positive correlation between two methods for DNase activity was observed: P lt 0.001 and Pearson correlation coefficient 0.740. Decreased DNase activity was found in 25 of 31 SLE patients (81%) by fluoresence-based method and in 24 of 31 SLE patients (77%) by ELISA test. We also observed the significant positive correlation between titer of anti-dsDNA antibodies and DNase activity measured by both methods (P lt 0.05). ConclusionsThe key improvement is the use of internal control in the fluorescence-based method, which diminishes the influence of technical errors on the obtained results and increases reliability of the assay. This improved fluorescence-based method, with additional validation, may provide an alternative to more expensive and time-consuming conventional methods, such as ELISA

DNA and biochemical analysis of a potential opaque2 maize population

Maize has low nutritional value because it is poor in essential amino acids lysine and tryptophan, but different mutations have been identified that increase their content. Two high lysine/tryptophan populations from Maize Research Institute genebank (IP1 and IP2) were identified in a previous research. In both populations, analysis with umc1066 opaque2 specific marker detected a recessive (o2), a dominant (O2) and an unknown allele (UA). However, IP2 lacked homozygous recessive o2o2 genotypes. The aim of the present research was to determine by DNA and biochemical analysis if UA allele was a recessive allele and/or if high tryptophan content was due to the o2 or some other mutation. Tree more opaque accessions with different mutations - IP3o5, IP4o14 and IP5floury (no data on type of mutation) were used in biochemical analysis for comparison with IP1 and IP2. Kernels were divided into two samples - with hard and with soft kernels. The UA allele sequencing revealed that it was a dominant allele with four GCCAGA repeats. SSR analysis showed presence of o2 in IP1 in both hard and soft kernels. Decrease in 22 kDa, 19 kDa and 27 kDa zeins in soft kernels was observed only in IP1 and IP2. Tryptophan content was high in soft kernels of IP1 (0.081) and IP2 (0.085), and in both hard and soft kernels of -IP3o5 (0.083 and 0.085, respectively). It can be concluded that IP1 is an o2 mutant and that IP2 carries a high tryptophan mutation other than o2, o5, o14 or floury

Comparative study of gamma and neutron irradiation effects on the silicon solar cells parameters

Zbog širokog područja primene, solarne ćelije su, u svom radnom okruženju, izložene različitim vrstama zračenja (kosmičko zračenje u gornjim slojevima atmosfere, vojna i civilna nuklearna postrojenja). Štaviše, korišćeno nuklearno gorivo u isto vreme emituje i u fotone i neutrone tako da se u solarnim ćelijama smeštenim u blizini ovih goriva javljaju različite vrste radijacionih oštećenja. Ova oštećenja su uzrokovana i gama i neutronskim zračenjem. Zbog toga su preduzimana vrlo opsežna istraživanja sa ciljem razvoja poluprovodničkih uređaja čiji rad će biti pouzdan i u uslovima povećanog nivoa zračenja. Sa tehnološkog aspekta, važno je utvrditi promene, uzrokovane zračenjem, u parametrima solarnih ćelija koje utiču na njihov rad. Cilj ovog rada je da se predstavi uporedna studija efekata gama i neutronskog ozračivanja na parametre solarnih ćelija.Due to its wide application areas, solar cells are exposed, in their work environment, to different types of radiation (cosmic radiation in the upper layers of the atmosphere, military and civilian nuclear facilities). Moreover, the used nuclear fuel emits y photons and neutrons at the same time, so different types of radiation damage appeared in solar cells located in the vicinity of these fuels. These damage have been caused by both gamma and neutron radiation. That's why very extensive researches have been undertaken with the aim of developing semiconductor devices whose work will be reliable in terms of increased levels of radiation. From the technological point of view, it is important to determine changes, caused by radiation, in the parameters of the silicon solar cells that affect their work. The aim of this paper is to present the comparative study of gamma and neutron irradiation effects on the solar cells parameters

The Impact of Successive Gamma and Neutron Irradiation on Characteristics of PIN Photodiodes and Phototransistors

The aim of this paper is to explore the impact of increased gamma and neutron radiation on the PIN photodiodes and phototransistors and their output characteristics. Special attention was paid to the successive impact of gamma and neutron radiation when the components were located in the field of gamma radiation and after that in the field of neutron radiation. The impact of successive irradiation was compared with the influence of gamma and neutron radiation when they appear individually. An important result of this research is the observation that neutron irradiation of photovoltaic detectors, applied after gamma irradiation, leading to partial reparations of distorted semiconductor structure and increasing disrupted output characteristics (photocurrent, spectral response). Monte Carlo simulation of gamma photons transfer through the crystal lattice of the semiconductor has been shown that the cause of such effect of neutron radiation is a large number of divacancies caused by successive operation of the previous gamma radiation and the neutron radiation itself. Divacancies have created the basis for increased generation of charge carriers by direct transfer (tunneling) of carriers through the traps (recombination centers). This is so called intercenter charge transfer

Comparative study of gamma and neutron irradiation effects on the silicon solar cells parameters

Zbog širokog područja primene, solarne ćelije su, u svom radnom okruženju, izložene različitim vrstama zračenja (kosmičko zračenje u gornjim slojevima atmosfere, vojna i civilna nuklearna postrojenja). Štaviše, korišćeno nuklearno gorivo u isto vreme emituje i u fotone i neutrone tako da se u solarnim ćelijama smeštenim u blizini ovih goriva javljaju različite vrste radijacionih oštećenja. Ova oštećenja su uzrokovana i gama i neutronskim zračenjem. Zbog toga su preduzimana vrlo opsežna istraživanja sa ciljem razvoja poluprovodničkih uređaja čiji rad će biti pouzdan i u uslovima povećanog nivoa zračenja. Sa tehnološkog aspekta, važno je utvrditi promene, uzrokovane zračenjem, u parametrima solarnih ćelija koje utiču na njihov rad. Cilj ovog rada je da se predstavi uporedna studija efekata gama i neutronskog ozračivanja na parametre solarnih ćelija.Due to its wide application areas, solar cells are exposed, in their work environment, to different types of radiation (cosmic radiation in the upper layers of the atmosphere, military and civilian nuclear facilities). Moreover, the used nuclear fuel emits y photons and neutrons at the same time, so different types of radiation damage appeared in solar cells located in the vicinity of these fuels. These damage have been caused by both gamma and neutron radiation. That's why very extensive researches have been undertaken with the aim of developing semiconductor devices whose work will be reliable in terms of increased levels of radiation. From the technological point of view, it is important to determine changes, caused by radiation, in the parameters of the silicon solar cells that affect their work. The aim of this paper is to present the comparative study of gamma and neutron irradiation effects on the solar cells parameters

ANALYSIS OF BUSINESS ENVIRONMENT USING THE MULTI-CRITERIA APPROACH – CASE OF BALKAN’S TRANSITION ECONOMIES

Due to the lack of their own financial resources, attracting the foreign direct investment (FDI) isthe main prerequisite for transitional economies in order to increase production and employment, sothat they can ensure the long term sustainable economic growth. In addition, the foreign directinvestment is an important instrument for the economy restructuring, based on market principles.However, achieving this goal is not simple at all. In order to attract foreign investors, it is necessaryto create a favorable business environment in transition countries, which requires a number ofeconomic, institutional, political and other reforms. The aim of this paper is to point out the mainfactors attracting foreign direct investment and, by using the multi-criteria approach, to rank theBalkan’s transition economies depending on the preferences of investors taking into account certaincomponents of the business environment

Inflammatory modulation of the response of bronchial epithelial cells to lipopolysaccharide with pretreatment by montelukast

Montelukast, a leukotriene receptor antagonist, is the most prescribed nonsteroidal anti-inflammatory drug used as an add-on therapy for asthma. Besides its effect on blocking leukotriene action, montelukast has been proposed to have secondary anti-inflammatory properties. This study aimed to investigate the modulatory effect of montelukast on the expression of major genes involved in airway inflammation (TNF, IL6) and remodeling (MMP9, TGFB1) in response to lipopolysaccharide (LPS) in vitro. The expression of selected genes was measured by quantitative real-time polymerase chain reaction 0h and 24h after LPS stimulation in cells pretreated with montelukast. Montelukast was found to significantly attenuate increased TNF and IL6 gene expression, to have a mild effect on MMP9 and have no effect on TGFB1 expression upon stimulation with LPS. The results of our study indicate that patients on montelukast therapy would have an adequate response to acute microorganism-induced inflammation, so additional anti-inflammatory effects of montelukast should be better exploited

Polyphenols impact on quality, safety and shelf life of fermented sausages

У раду је испитивана могућност употребе полифенола као природног конзерванса код ферментисаних кобасица. Произведене су три групе производа, од којих је прва била кобасица уобичајеног састава са додатком нитрита, друга група је била истог састава као прва с тим што су јој додати и полифеноли, а трећа је произведена без додатка нитрита али са додатком полифенола. Резултати испитивања су показали да додатак полифенола није утицао на процесе ферментације и сушења, а производи су били стандардног хемијског састава. Кобасице произведене са нитритима и полифенолима садржале су више нитрата него контрола. Код кобасица обогаћених полифенолима утврђен је значајно мањи пероксидни и TBARS број него код контроле. Међутим, код кобасица произведених заједно са нитритима и полифенолима утврђен је јачи интензитет оксидације масти него код кобасице са полифенолима без нитрита. Микробиолошки процеси код свих група били су типични за ферментисане кобасице, у погледу броја бактерија млечне киселине и фамилије Micrococcaceae. Кобасице обогаћене полифенолима садржале су мањи број бактерија узрочника квара (Pseudomonadaceae) него контролне кобасице. Иако су инструменталним мерењем регистроване разлике у светлоћи, уделу црвене и жуте боје, сензорска својства свих група кобасица су високо оцењена током већег дела складиштења, а 280. дана биле су у оквиру лимита прихватљивости. Кобасице са полифенолима садржале су више биогених амина него контролне кобасице, при чему су кобасице без нирита садржале више биогених амина него кобасице са нитритима. Садржај хистамина и тирамина био је у границама уобичајеним за ферментисане кобасице. Најдоминантнија фенолна једињења била су каемферол-3-О-глукозид, а затим и кверцетин, лутеолин-7-О-глукозид, катехин и сирингинска киселина. Кобасице са полифенолима без додатка нитрита достигле су исти период одрживости као конвенционалне кобасице са нитритима, што је обећавајући резултат у потенцијалној употреби полифенола као замене за нитрите, при чему је потребно пронаћи адекватно решење како би се смањило стварање биогених амина.In this work, the possibility to use polyphenols as natural preservatives in fermented sausage was investigated. Three groups of sausages were produced, the first group were sausages of usual composition containing nitrite, the second group were sausages of the same composition as the first but with the addition of a polyphenols, and the third were sausages produced without the addition of nitrites in which were added polyphenols. The results showed that the addition of polyphenols did not affect the processes of fermentation and drying and the final products had the usual chemical composition. The sausages produced with nitrites and polyphenols contained higher nitrate levels than the control. By the polyphenols enriched sausages there was found a significantly lower peroxide number and TBARS value than by the control sausage. However, for sausages manufactured together with nitrites and polyphenols, a higher intensity of fat oxidation processes was found then by nitrite free sausages. Although the instrumental measurements registered differences in brightness, red and yellow color intensity, the sensory properties of all groups of sausages were rated high for most of the storage period and reached a day 280 with scores within the limits of acceptability. Polyphenols enriched sausages contained more biogenic amines compared to the control sausages, while the sausages without nitrites contained more biogenic amines then nitrite containing sausages. Content of histamine and tiramine was in range usual for fermented sausges. The dominant phenolic compounds in sausages were kaemferol-3-O-glucoside, quercetin, and then, luteolin-7-O-gukozid, catechin and syringic acid. Sausages with polyphenols produced without nitrites reached the same shelf life as the conventional nitrite containing sausages, which is a promising result in the potential use of polyphenols as a substitute for nitrite in fermented sausages, but it is necessary to find an adequate solution in order to reduce the biogenic amines formation in these sausages