structural-information-from-single-molecule-fret-experiments-using

Structural Information from Single-molecule FRET Experiments Using the Fast Nano-positioning System

Single-molecule F&#246;rster Resonance Energy Transfer (smFRET) can be used to obtain structural information on biomolecular complexes in real-time. ...

Ulm University

Proteoglycans play key roles in the physiological assembly of extracellular matrices and in the modulation of growth factor activities. During liver regeneration there is a profound remodelling of the connective tissue network with a concurrent alteration in proteoglycan gene expression. In the present study we have analyzed in detail the biochemical and molecular properties of the proteoglycans associated with biliary cirrhosis. The three major proteoglycans of human liver, namely decorin, syndecan and perlecan, were markedly elevated in the cirrhotic parenchyma as compared to normal liver tissue. Particularly elevated (eight fold) was the perlecan. This proteoglycan had not only heparan sulfate but also chondroitin and dermatan sulfate. Reverse transcriptase PCR revealed a marked enhancement of decorin and syndecan expression and detectable message for perlecan was found only in the cirrhotic liver. These results indicate that significant proteoglycan alterations are associated with the development of biliary cirrhosis and provide basis for future studies aimed at the characterization of the molecular events involved in the regulation of extracellular matrix deposition in this common human disease.

Altered Proteoglycan Gene Expression in Human Biliary Cirrhosis.

In this paper we give a practical overview of the changes in the financing of health care in Hungary. We describe the financing system of general practitioners, home care (nursing), out-patient care and the acute and chronic care of hospitals. We show how the financial system has changed after the political changes of 1990. The global budget approach of the 1980s was replaced by performance-related financing methods including the ICPM (International Classification of Procedures in Medicine) code system of the WHO (World Health Organization) in out-patient care and the introduction of HBCS (Homogen Betegsegcsoportok, "Homogeneous Disease Groups") in in-patient care. We underline that the efforts made towards reforming health care financing resulted in an activity-related financing system.

Financing of health care services in Hungary.

On the basis of documentary analysis and interviews with decision makers, this paper discusses the cost accounting methodologies used for price setting of inpatient services in the Hungarian health care system focusing on sector of acute inpatient care, which is financed through the Hungarian adaptation of Diagnosis Related Groups since 1993. Hungary has a quite sophisticated DRG system, which had a deep impact on the efficiency of the acute inpatient care sector. Nevertheless, the system requires continuous maintenance, where the cooperation of hospitals, as well as the minimisation of political influence are critical success factors.

Cost accounting methodologies in price setting of acute inpatient services in Hungary.

TFIIE and the archaeal homolog TFE enhance DNA strand separation of eukaryotic RNAPII and the archaeal RNAP during transcription initiation by an unknown mechanism. We have developed a fluorescently labeled recombinant M. jannaschii RNAP system to probe the archaeal transcription initiation complex, consisting of promoter DNA, TBP, TFB, TFE, and RNAP. We have localized the position of the TFE winged helix (WH) and Zinc ribbon (ZR) domains on the RNAP using single-molecule FRET. The interaction sites of the TFE WH domain and the transcription elongation factor Spt4/5 overlap, and both factors compete for RNAP binding. Binding of Spt4/5 to RNAP represses promoter-directed transcription in the absence of TFE, which alleviates this effect by displacing Spt4/5 from RNAP. During elongation, Spt4/5 can displace TFE from the RNAP elongation complex and stimulate processivity. Our results identify the RNAP "clamp" region as a regulatory hot spot for both transcription initiation and transcription elongation.

The initiation factor TFE and the elongation factor Spt4/5 compete for the RNAP clamp during transcription initiation and elongation.

In recent times, the topic of smoking has been extensively debated in Hungary. A new Act has been issued for the protection of non-smokers and for the regulation of tobacco product distribution.

[Health related costs of smoking from the viewpoint of the Health Insurance Fund in Hungary].

The molecular architecture of RNAP II-like transcription initiation complexes remains opaque due to its conformational flexibility and size. Here we report the three-dimensional architecture of the complete open complex (OC) composed of the promoter DNA, TATA box-binding protein (TBP), transcription factor B (TFB), transcription factor E (TFE) and the 12-subunit RNA polymerase (RNAP) from Methanocaldococcus jannaschii. By combining single-molecule Förster resonance energy transfer and the Bayesian parameter estimation-based Nano-Positioning System analysis, we model the entire archaeal OC, which elucidates the path of the non-template DNA (ntDNA) strand and interaction sites of the transcription factors with the RNAP. Compared with models of the eukaryotic OC, the TATA DNA region with TBP and TFB is positioned closer to the surface of the RNAP, likely providing the mechanism by which DNA melting can occur in a minimal factor configuration, without the dedicated translocase/helicase encoding factor TFIIH.

Complete architecture of the archaeal RNA polymerase open complex from single-molecule FRET and NPS.

Cannabimimetic compounds have gained an increasing attention from the forensic community during the past few years. The present study was aimed to develop a liquid chromatographic separation method for the analysis of JWH-122 and its methyl isomers. In Hungary, JWH-122 is scheduled as a narcotic compound and its methyl isomers fall into the new psychoactive substance category, attracting significantly milder punishment than JWH-122 does. According to our best knowledge, gas chromatography or reversed phase liquid chromatography coupled with mass spectrometry could not be applied for separation and selective determination of methyl-naphthoyl indol isomers. In this study, we aimed to develop a high performance liquid chromatography method with UV and mass spectrometric detection for the separation of JWH-122 and all its possible isomers, depending on the position of methyl group on the naphthyl frame. Different reversed phase columns were used. Alkyl-modified silica with different selectivity and morphology with different mobile phase composition cannot be applied for separation of JWH-122 isomers. Porous graphitic carbon (PGC) column was used for separation of banned JWH-122 and each of its methyl isomers. In method development, a Quality by Design approach is presented for modeling the retention of the compounds. According to our knowledge, this is the first time reporting the use of intelligent software to estimate the retention on PGC material and using non-aqueous conditions. Retention times predicted by two program packages (STATISTICA(®) and DryLab(®)) are compared. The possibilities and limitations of the software modeling in the conditions described above are also evaluated.

Separation of Isomers of JWH-122 on Porous Graphitic Carbon Stationary Phase with Non-Aqueous Mobile Phase Using Intelligent Software.

Angioedema is a rare and serious adverse drug reaction (ADR) to angiotensin-converting enzyme (ACE) inhibitor treatment. Dry cough is a common side effect of ACE inhibitors and has been identified as a possible risk factor for angioedema.

Julia Nagy

Institute of Biophysics

Ulm University

Structural Information from Single-molecule FRET Experiments Using the Fast Nano-positioning System

Julia Nagy

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Institute of Biophysics

Ulm University

Structural Information from Single-molecule FRET Experiments Using the Fast Nano-positioning System

Institute of Biophysics