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Search Results: 1 - 10 of 121355 matches for " Szilvia Tóth "
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Restriction Enzyme Body Doubles and PCR Cloning: On the General Use of Type IIS Restriction Enzymes for Cloning
Eszter Tóth, Krisztina Huszár, Petra Bencsura, Péter István Kulcsár, Barbara Vodicska, Antal Nyeste, Zsombor Welker, Szilvia Tóth, Ervin Welker
PLOS ONE , 2014, DOI: 10.1371/journal.pone.0090896
Abstract: The procedure described here allows the cloning of PCR fragments containing a recognition site of the restriction endonuclease (Type IIP) used for cloning in the sequence of the insert. A Type IIS endonuclease - a Body Double of the Type IIP enzyme - is used to generate the same protruding palindrome. Thus, the insert can be cloned to the Type IIP site of the vector without digesting the PCR product with the same Type IIP enzyme. We achieve this by incorporating the recognition site of a Type IIS restriction enzyme that cleaves the DNA outside of its recognition site in the PCR primer in such a way that the cutting positions straddle the desired overhang sequence. Digestion of the PCR product by the Body Double generates the required overhang. Hitherto the use of Type IIS restriction enzymes in cloning reactions has only been used for special applications, the approach presented here makes Type IIS enzymes as useful as Type IIP enzymes for general cloning purposes. To assist in finding Body Double enzymes, we summarised the available Type IIS enzymes which are potentially useful for Body Double cloning and created an online program (http://group.szbk.u-szeged.hu/welkergr/b?ody_double/index.html) for the selection of suitable Body Double enzymes and the design of the appropriate primers.
Exposure to Lipopolysaccharide and/or Unconjugated Bilirubin Impair the Integrity and Function of Brain Microvascular Endothelial Cells
Filipa L. Cardoso, ágnes Kittel, Szilvia Veszelka, Inês Palmela, Andrea Tóth, Dora Brites, Mária A. Deli, Maria A. Brito
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0035919
Abstract: Background Sepsis and jaundice are common conditions in newborns that can lead to brain damage. Though lipopolysaccharide (LPS) is known to alter the integrity of the blood-brain barrier (BBB), little is known on the effects of unconjugated bilirubin (UCB) and even less on the joint effects of UCB and LPS on brain microvascular endothelial cells (BMEC). Methodology/Principal Findings Monolayers of primary rat BMEC were treated with 1 μg/ml LPS and/or 50 μM UCB, in the presence of 100 μM human serum albumin, for 4 or 24 h. Co-cultures of BMEC with astroglial cells, a more complex BBB model, were used in selected experiments. LPS led to apoptosis and UCB induced both apoptotic and necrotic-like cell death. LPS and UCB led to inhibition of P-glycoprotein and activation of matrix metalloproteinases-2 and -9 in mono-cultures. Transmission electron microscopy evidenced apoptotic bodies, as well as damaged mitochondria and rough endoplasmic reticulum in BMEC by either insult. Shorter cell contacts and increased caveolae-like invaginations were noticeable in LPS-treated cells and loss of intercellular junctions was observed upon treatment with UCB. Both compounds triggered impairment of endothelial permeability and transendothelial electrical resistance both in mono- and co-cultures. The functional changes were confirmed by alterations in immunostaining for junctional proteins β-catenin, ZO-1 and claudin-5. Enlargement of intercellular spaces, and redistribution of junctional proteins were found in BMEC after exposure to LPS and UCB. Conclusions LPS and/or UCB exert direct toxic effects on BMEC, with distinct temporal profiles and mechanisms of action. Therefore, the impairment of brain endothelial integrity upon exposure to these neurotoxins may favor their access to the brain, thus increasing the risk of injury and requiring adequate clinical management of sepsis and jaundice in the neonatal period.
Nesfatin-1/NUCB2 as a Potential New Element of Sleep Regulation in Rats
Szilvia Vas, Csaba ádori, Katalin K?ncz?l, Zita Kátai, Dorottya Pap, Rege S. Papp, Gy?rgy Bagdy, Miklós Palkovits, Zsuzsanna E. Tóth
PLOS ONE , 2013, DOI: 10.1371/journal.pone.0059809
Abstract: Study Objectives Millions suffer from sleep disorders that often accompany severe illnesses such as major depression; a leading psychiatric disorder characterized by appetite and rapid eye movement sleep (REMS) abnormalities. Melanin-concentrating hormone (MCH) and nesfatin-1/NUCB2 (nesfatin) are strongly co - expressed in the hypothalamus and are involved both in food intake regulation and depression. Since MCH was recognized earlier as a hypnogenic factor, we analyzed the potential role of nesfatin on vigilance. Design We subjected rats to a 72 h-long REMS deprivation using the classic flower pot method, followed by a 3 h-long ‘rebound sleep’. Nesfatin mRNA and protein expressions as well as neuronal activity (Fos) were measured by quantitative in situ hybridization technique, ELISA and immunohistochemistry, respectively, in ‘deprived’ and ‘rebound’ groups, relative to controls sacrificed at the same time. We also analyzed electroencephalogram of rats treated by intracerebroventricularly administered nesfatin-1, or saline. Results REMS deprivation downregulated the expression of nesfatin (mRNA and protein), however, enhanced REMS during ‘rebound’ reversed this to control levels. Additionally, increased transcriptional activity (Fos) was demonstrated in nesfatin neurons during ‘rebound’. Centrally administered nesfatin-1 at light on reduced REMS and intermediate stage of sleep, while increased passive wake for several hours and also caused a short-term increase in light slow wave sleep. Conclusions The data designate nesfatin as a potential new factor in sleep regulation, which fact can also be relevant in the better understanding of the role of nesfatin in the pathomechanism of depression.
Edaravone Protects against Methylglyoxal-Induced Barrier Damage in Human Brain Endothelial Cells
Andrea E. Tóth, Fruzsina R. Walter, Alexandra Bocsik, Petra Sántha, Szilvia Veszelka, Lajos Nagy, László G. Puskás, Pierre-Olivier Couraud, Fuyuko Takata, Shinya Dohgu, Yasufumi Kataoka, Mária A. Deli
PLOS ONE , 2014, DOI: 10.1371/journal.pone.0100152
Abstract: Background Elevated level of reactive carbonyl species, such as methylglyoxal, triggers carbonyl stress and activates a series of inflammatory responses leading to accelerated vascular damage. Edaravone is the active substance of a Japanese medicine, which aids neurological recovery following acute brain ischemia and subsequent cerebral infarction. Our aim was to test whether edaravone can exert a protective effect on the barrier properties of human brain endothelial cells (hCMEC/D3 cell line) treated with methylglyoxal. Methodology Cell viability was monitored in real-time by impedance-based cell electronic sensing. The barrier function of the monolayer was characterized by measurement of resistance and flux of permeability markers, and visualized by immunohistochemistry for claudin-5 and β-catenin. Cell morphology was also examined by holographic phase imaging. Principal Findings Methylglyoxal exerted a time- and dose-dependent toxicity on cultured human brain endothelial cells: a concentration of 600 μM resulted in about 50% toxicity, significantly reduced the integrity and increased the permeability of the barrier. The cell morphology also changed dramatically: the area of cells decreased, their optical height significantly increased. Edaravone (3 mM) provided a complete protection against the toxic effect of methylglyoxal. Co-administration of edaravone restored cell viability, barrier integrity and functions of brain endothelial cells. Similar protection was obtained by the well-known antiglycating molecule, aminoguanidine, our reference compound. Conclusion These results indicate for the first time that edaravone is protective in carbonyl stress induced barrier damage. Our data may contribute to the development of compounds to treat brain endothelial dysfunction in carbonyl stress related diseases.
A New Zearalenone Biodegradation Strategy Using Non-Pathogenic Rhodococcus pyridinivorans K408 Strain
Rókus Kriszt, Csilla Krifaton, Sándor Szoboszlay, Mátyás Cserháti, Balázs Kriszt, József Kukolya, árpád Czéh, Szilvia Fehér-Tóth, Lívia T?r?k, Zsuzsanna Sz?ke, Krisztina J. Kovács, Teréz Barna, Szilamér Ferenczi
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0043608
Abstract: Zearalenone (hereafter referred to as ZEA) is a nonsteroidal estrogenic mycotoxin produced by several Fusarium spp. on cereal grains. ZEA is one of the most hazardous natural endocrine disrupting chemicals (EDC) which induces hyper estrogenic responses in mammals. This can result in reproductive disorders in farm animals as well as in humans. Consequently, detoxification strategies for contaminated crops are crucial for food safety. In this study we have developed a bacterial based detoxification system using a non-pathogen Rhodococcus pyridinivorans K408 strain. Following 5 days treatment of ZEA with R. pyridinivorans K408 strain HPLC analyses showed an 87.21% ZEA-degradation efficiency of the bacterial enzyme systems. In another approach, the strain biotransformation ability has also been confirmed by a bioluminescent version of the yeast estrogen screening system (BLYES), which detected an 81.75% of biodegradability of ZEA, in a good agreement with the chemical analyses. Furthermore, the capacity of R. pyridinivorans to eliminate the estrogenic effects of ZEA was tested by using an immature uterotrophic assay. Prepubertal female rats were treated with vehicle (olive oil), 17β-estradiol, ZEA (0.1-1-5-10 mg/kg body weight) and LB broth containing 500 mg/l ZEA that has already been incubated with or without Rhodococcus pyridinivorans K408 strain. Uterine weights were measured and the mRNA level changes relating to apelin, aquaporin 5, complement component 2, and calbindin-3 genes were measured by qRT-PCR. These genes represent the major pathways that are affected by estromimetic compounds. Zearalenone feeding significantly increased the uterus weight in a dose dependent manner and at the same time upregulated complement component 2 and calbindin-3 expression as well as decreased apelin and aquaporin 5 mRNA levels comparable to that seen in 17β-estradiol exposed rats. In contrast, LB broth in which ZEA was incubated with Rhodococcus pyridinivorans K408 prior to the feeding did not display any estrogenic effect neither on uterine weight nor on the expression of estrogen-regulated genes. Consequently, the identification of Rhodococcus pyridinivorans K408 strain in ZEA biodegradation proved to be a very efficient biological tool that is able to eliminate the complete estrogenic effects of ZEA. It is also remarkable that this biotransformation pathway of ZEA did not result in any residual estrogenic effects.
Erythropoietin Receptor Expression Is a Potential Prognostic Factor in Human Lung Adenocarcinoma
Anita Rózsás, Judit Berta, Lívia Rojkó, László Z. Horváth, Magdolna Keszthelyi, István Kenessey, Viktória László, Walter Berger, Michael Grusch, Mir Alireza Hoda, Szilvia T?r?k, Walter Klepetko, Ferenc Rényi-Vámos, Balázs Heged?s, Balázs D?me, József Tóvári
PLOS ONE , 2013, DOI: 10.1371/journal.pone.0077459
Abstract: Recombinant human erythropoietins (rHuEPOs) are used to treat cancer-related anemia. Recent preclinical studies and clinical trials, however, have raised concerns about the potential tumor-promoting effects of these drugs. Because the clinical significance of erythropoietin receptor (EPOR) signaling in human non-small cell lung cancer (NSCLC) also remains controversial, our aim was to study whether EPO treatment modifies tumor growth and if EPOR expression has an impact on the clinical behavior of this malignancy. A total of 43 patients with stage III–IV adenocarcinoma (ADC) and complete clinicopathological data were included. EPOR expression in human ADC samples and cell lines was measured by quantitative real-time polymerase chain reaction. Effects of exogenous rHuEPOα were studied on human lung ADC cell lines in vitro. In vivo growth of human ADC xenografts treated with rHuEPOα with or without chemotherapy was also assessed. In vivo tumor and endothelial cell (EC) proliferation was determined by 5-bromo-2’-deoxy-uridine (BrdU) incorporation and immunofluorescent labeling. Although EPOR mRNA was expressed in all of the three investigated ADC cell lines, rHuEPOα treatment (either alone or in combination with gemcitabine) did not alter ADC cell proliferation in vitro. However, rHuEPOα significantly decreased tumor cell proliferation and growth of human H1975 lung ADC xenografts. At the same time, rHuEPOα treatment of H1975 tumors resulted in accelerated tumor endothelial cell proliferation. Moreover, in patients with advanced stage lung ADC, high intratumoral EPOR mRNA levels were associated with significantly increased overall survival. This study reveals high EPOR level as a potential novel positive prognostic marker in human lung ADC.
A mosás és tisztaság kultúrt rténete Budapesten a két világháború k z tt. A mosón k korán halnak… The washing and cleanliness cultural history of Budapest, between the two World Wars. The laundresses die early…
Czingel Szilvia
Kaleidoscope History , 2013,
Abstract:
Necessity of Next Generation Network Infrastructure and the Evaluation of Broadband Developments in Rural Regions
Szilvia Botos
Agrárinformatika Folyóirat , 2012,
Abstract: Impacts of next generation network for the condition and development of rural regions are already evident, but the different economic sectors be differently affected by the effects of ICT and they require it in different degrees. In the national and EU strategies, building next generation infrastructure in rural areas, by this decreasing the extent of digital gap and reducing the disparities, is one of the main goals. We have proved by regional analysis that the infrastructural and usage features, which link to broadband networks (e.g. households with broadband access, shopping through Internet or the rate of computer usage), these be in connected with the socio-economic features, like regional production of GDP or the employment rate. Based on results, we could determine that the differences which be showed in the ICT level between the countries, they present also between the regions, within one country. For analyzing the levels of development of the countries, several indicators are available and these help to determine the future development directions, and some of those we will show in this article. But first, we give some review about the significance of NGN and e-services in the less favoured regions, the barriers of improvements and the comparison possibilities of the countries relative to their ICT situation in different territorial levels.
FOREIGN LANGUAGE LEARNING AND EDUCATION IN MINORITY CIRCUMSTANCES. PROBLEMS AND POSSIBLE SOLUTIONS
Szilvia Batyi
Eesti ja soome-ugri keeleteaduse ajakiri , 2011,
Abstract: Some form of bi- and multilingualism means the natural lingual condition for more than the half of the population of the Earth. It is a substantial linguistic aim of the Transcarpathian Hungarian community that beside preservation of their mother tongue(the Hungarian), acquire the state language (the Ukrainian language)and the basis of at least one world language. But this aim is hindered by a lot of things in Transcarpathia. The goal of the study is to shed light on these problems and to find possible solutions based on two researches. The first research, which was carried out in the Tanscarpathian Hungarian schools, was to reveal the conditions and problems of foreign language education. The research threw light on numerous problems that approve the low level of foreign language knowledge of the Transcarpathian Hungarian youth. Attitudes and stereotypes influence the success of foreign language acquisition. For this reason in the second part of the study I would like to show, what kind of stereotypes and attitudes can be discovered in the parents (who are lay linguistically and language pedagogically)concerning foreign languages, and within this especially concerning the English language. It appears from the interviews,that nor the attitudes of the state towards foreign languages that was inherited from the soviet system, neither the impassiveness of the parents improves the positive attitudes in the Transcarpathian Hungarian students towards foreign languages, and nor the state,neither the parents approve the motivation of foreign language acquisition.
Walther Kindt: Irrtümer und andere Defizite in der Linguistik. Wissenschaftslogische Probleme als Hindernis für Erkenntnisfortschritte
Tóth,té
Argumentum , 2012,
Abstract: Review
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