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Acta Biologica Hungarica
Authors:
Nóra Faragó
,
Ágnes Zvara
,
Z. Varga
,
P. Ferdinandy
, and
L. Puskás

Micro RNAs (miRNA) are an abundant class of small RNAs that regulate the stability and translation of cognate mRNAs. MiRNAs are potential diagnostic markers, moreover, they play an essential role in the development of various heart disesases. In case of limited tissue material, such as, e.g. human biopsies, purification of miRNAs with sufficient yield is critical. Reproducible expression analysis of miRNAs is highly dependent on the quality of the RNA, which is often difficult to achieve from fibrous tissue such as the heart. Several companies developed general purification kits for miRNAs, however, none of them are specialized to fibrotic tissues. Here we describe an optimized miRNA purification protocol that results in high miRNA yield as compared to other methods including trizol-based and column-based protocols. By using our improved protocol, miRNA obtained from heart tissue gave more reproducible results in QRT-PCR analysis and obtained more significant calls (172 vs. 118) during DNA microarray analysis when compared to the commercially available kit. In addition to the heart tissue, the present protocol can be applied to other fibrotic tissues, such as lung or skeletal muscle to isolate high-purity miRNA.

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The purpose of this work was to determine the individual and combined effects of insecticide Sumithion 50 EC (50% fenitrothion) and herbicide Fusilade S (12.5% fluazifop-P-butyl) on the development of pheasant embryos. Eggs were treated by injection of various concentrations of pesticides into the air space on day 12 of incubation. Pathological examination of embryos was carried out on day 23 of the hatching period. Mortality rate, body weight data and morphological alterations were evaluated after the macroscopic examination. The skeletal staining method was used to detect deformities. The two pesticides used in combination moderated the toxic/teratogenic effects of individual treatment.

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Toxicological studies on wild animals play an important role in the ecotoxicological examination of pesticides. The applied model tests enable the assessment of toxicological consequences with particular regard to the life and nutrition of wild animals in the ploughed field among plants treated with pesticides. The application of different pesticide formulations on plough-land may pose a simultaneous chemical burden to wild birds. In this model study, manifestations of the interaction between an insecticide and a herbicide were studied in pheasants. The birds were placed on lucerne in cages (48 m2) and sprayed once. The applied doses were: Sumithion 50 EC 1 litre/ha + Fusilade S 6 litres/ha (practical doses) and Sumithion 50 EC 5 litres/ha + Fusilade S 30 litres/ha. The analytically determined pesticide concentration of the lucerne was taken as a basis in the further treatment of fodder. The fodder of pheasants contained the following chemicals: 85 mg/kg Sumithion 50 EC + 510 mg/kg Fusilade S and 425 mg/kg Sumithion 50 EC + 2250 mg/kg Fusilade S. Sporadic deaths observed among the pheasants were of traumatic origin and not due to a toxic effect. The decrease of body weight was significant only at the higher dose levels. Acetylcholinesterase (AChE) activity of the blood decreased significantly in both dose groups. On the basis of the results obtained it can be established that at the dose level used in the practice the pesticides studied do not give rise to a toxic interaction in pheasants.

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In order to improve the isolation rate of Rhodococcus equi from animals and soil, the efficacy of four previously described selective media (CAZ-NB, M3T, NANAT and TINSDALE) and that of four other media (NC, PNP, TCP and TVP) composed by us was compared and evaluated. Two selective plating media proved to be the best for the isolation of R. equi from contaminated samples. One of them was CAZ-NB containing ceftazidime, novobiocin and cycloheximide, while the other was the newly composed TCP containing trimethoprim, cefoperazone, polymyxin B, cycloheximide and potassium tellurite as selective components. These two media allowed the growth of at least 62-72% of R. equi present in the artificially contaminated samples, and the inhibition of unwanted contaminant bacteria and fungi was satisfactory with both media. TCP medium proved to be superior to CAZ-NB since the colony morphology of R. equi was much more characteristic (shiny, smooth, black colonies 3-5 mm in diameter) on it, and it inhibited the unwanted contaminant bacterial and fungal flora more effectively, especially in the case of faecal and soil samples. Therefore, TCP is recommended as a new, highly selective plating medium for the isolation of R. equi from contaminated samples.

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Ground ivy (Glechoma hederacea L.) is one of the prosperous plants for the food-industry as natural antioxidant. This fact led us to examine the chemical diversity of six ground ivy populations situated in different natural habitats and to analyse the effect of the harvesting time. Total phenolic content, chlorogenic acid, and rutin content, as well as the antioxidant capacity showed significant differences due to the harvest time. The highest total phenol content (115 mg g–1 GAE) and the strongest antioxidant activity (53.3 mg g–1 AAE) were measured in the population originated from Budapest (GLE 6), harvested in July. The highest chlorogenic acid (357 mg/100 g) and rutin (950 mg/100 g) contents were detected in the July harvested samples from the Soroksár Botanical Garden population (GLE 1). According to our results, the collection time has significant effect on the total phenolic content – first of all on the chlorogenic acid and rutin accumulation levels of ground ivy, while the influence of the habitat seems to be less important.

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Journal of Radioanalytical and Nuclear Chemistry
Authors:
G. Duplâtre
,
Zs. Kajcsos
,
T. Goworek
,
L. Varga
,
L. Liszkay
,
I. Billard
, and
K. Lázár

Abstract  

Positron annihilatioin data collected for zeosil (a zeoilite-type solid allowing abundant Psformation) are discussed, with particular attention payed to the longest-lived positronium states as observed in lifetime spectroscopy experiments. To unravel the nature of these components, additional information was sought by applying an extermal magnetic field and by reecording the total energy distributioin of the annihilation radiation. From both the lifetime and the extended Doppler spectra, it is shown that positronium presents a very long-lived component with a relative intensity of ca. 30% and a lifetime close to the intrinsic tripler lifetime (140ns), therefore undergoing almost no pick-off annihilation. From the magnetic field experiments, the contact density parameter has a value of essentially 1, which is characteristic of an unpoerturbed triplet Ps state in vacuo. This positronium state therefore does not appear to interact with the medium.

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Acta Alimentaria
Authors:
M. Szalai
,
J. Szigeti
,
L. Farkas
,
L. Varga
,
A. Réti
, and
E. Zukál

The shelf-lives of major commercial cooked meat products (i.e., Bologna sausage, Italian-type cooked sausage, and cooked ham) packaged under vacuum or modified atmospheres were tested in this study. Samples were taken from commercial meat processing lines, sliced to 1.2 mm thickness and placed overlapped into polypropylene trays sealed with plastic films. The headspace of modified atmosphere packaged formulations consisted of 30% CO2 and 70% N2 or 60% CO2 and 40% N2. The samples thus produced were stored under refrigerated conditions. The values of microbiological, chemical, physical or sensory properties were plotted against storage time, and Gompertz curves were fitted to all time series that changed from an initial to a final value during any period of storage. The influence of headspace CO2 concentration on the properties of sliced cooked meat products varied considerably and, therefore, it was not possible to specify general rules. However, the presence of CO2 in the packaging atmosphere slowed down the rate of microbial growth, thereby delaying the spoilage of meat products. A CO2 level of 60% had beneficial effects on both the microbiological and sensory properties of sliced sausages and cooked ham. It was concluded that cooked meat products packaged under modified atmospheres had a shelf-life of 20 days.

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Acta Microbiologica et Immunologica Hungarica
Authors:
Ilona Dóczi
,
Erika Dósa
,
J. Varga
,
Zsuzsanna Antal
,
L. Kredics
, and
Elizabeth Nagy

The purpose of this study was to evaluate the Etest as an in vitro antifungal susceptibility test method for different moulds originating from human samples and from the environment. A total of 50 isolates (1 Acremonium, 18 Aspergillus, 2 Cladosporium, 1 Epicoccum, 15 Penicillium, 2 Scopulariopsis and 11 Trichoderma strains) were tested by the Etest. Forty-six of the tested moulds (92%) were resistant to fluconazole with minimal inhibitory concentrations (MICs) ³ 256 µg ml-1. There were strains resistant to ketoconazole among Aspergillus niger, A. ochraceus and Cladosporium spp. with MICs ? 32 µg ml-1. For fluconazole, no differences were observed using two different inocula, while for itraconazole, ketoconazole and amphotericin B, a 1 or less step 2-fold dilution difference in MIC was seen for the most of 10 selected strains. The MICs of fluconazole and amphotericin B obtained for Trichoderma strains by the Etest and the agar dilution method were also compared. MICs for fluconazole were in agreement, while MICs for amphotericin B were higher with 1 or 2 steps of 2-fold dilutions for most of Trichoderma strains in the case of the agar dilution method.

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The fungal revolution taking place in otorhinology inspired us to study the frequency of occurrence of fungi in the nasal mucus of chronic rhinosinusitis (CRS) patients (with or without polyposis) in order to evaluate the incidence of eosinophilic fungal sinusitis in CRS patients. Ninety-six samples were examined from patients with CRS. In 74 cases mucus was collected non-invasively, and in 22 cases during operation. The Gram-stained direct smears of all samples were also evaluated. Bacteria and fungi colonizing in the mucus were detected by culturing method. The control group consisted of 50 healthy volunteers. Typical aerobic pathogenic bacteria could be isolated from 34 patients. Fifty-seven aerobic bacteria were isolated, i.e. 1.6 bacteria/positive patient with a maximum of 3 different bacteria/sample. The most frequently isolated bacteria were Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Streptococcus pneumoniae, and Haemophilus influenzae. Yeasts and moulds could be detected from 79 patients (83%): Candida albicans, Candida spp., Aspergillus spp., Cladosporium spp, and Penicillium spp. were isolated most frequently. Altogether 237 yeasts and moulds were isolated, i.e. 3.0 different fungi/positive patient, with a maximum of 5 different fungi/sample. In the control group aerobic pathogens were not isolated, only apathogenic species. Fungi were isolated from 22 healthy patients (44%). These data indicate that fungi are frequently involved in the aetiology of CRS. IgE-mediated hypersensitivity to fungal allergens could not be proven in our patients.

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The aim of the present study was to identify gene expression changes in the rapid cardiac pacing-induced delayed antiarrhythmic protection in the canine, using cDNA microarrays and quantitative real-time PCR (QRT -PCR) techniques. In all dogs under light pentobarbitone anaesthesia, a pacing electrode was introduced into the right ventricle, and then the animals were divided into three groups: (1) sham-operated and sham-paced group (SP, n = 3) (2) ischaemic control group (IC; n = 3); these were without cardiac pacing and subjected only to a 25 min occlusion of the left anterior descending coronary artery (LAD), and (3) paced group (PC, n = 3); these animals were paced at a rate of 220–240 beats min−1 24 h prior to ischaemia. With cDNA chip 23 genes were found with altered expression in response to rapid cardiac pacing and 10 genes in the IC group when compared to SP dogs. These genes encode transcription factors (MEF2); members of signaling pathways (TGFβ2, PDE4D9), hormone related proteins (e.g. vasopressin V1 and V2 receptors). RT-QPCR was used either to confirm the results of the microarray analysis and also to study 46 genes which are already known to have a role in the late phase of PC. By this method 17 genes were up-regulated and 6 genes down-regulated in the IC group; their expression ratios changed either to the opposite or showed no alteration after cardiac pacing. This study would add some new information about those transcriptional changes that are involved in the delayed phase of cardiac protection.

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