Probing molecular pathways for DNA orientational trapping, unzipping and translocation in nanopores by using a tunable overhang sensor.

Nanopores provide a unique single-molecule platform for genetic and epigenetic detection. The target nucleic acids can be accurately analyzed by characterizing their specific electric fingerprints or signatures in the nanopore. Here we report a series of novel nanopore signatures generated by target nucleic acids that are hybridized with a probe. A length-tunable overhang appended to the probe functions as a sensor to specifically modulate the nanopore current profile. The resulting signatures can reveal multiple mechanisms for the orientational trapping, unzipping, escaping and translocation of nucleic acids in the nanopore. This universal approach can be used to program various molecular movement pathways, elucidate their kinetics, and enhance the sensitivity and specificity of the nanopore sensor for nucleic acid detection.

Differentiation and characterization in primary culture of white adipose tissue brown adipocyte-like cells.

BACKGROUND: Rodent brown adipose tissue (BAT) is considered the main effector of adaptative thermogenesis as it contains a unique mitochondrial uncoupling protein, termed as uncoupling protein-1 (UCP1). The emergence of ectopic brown adipocytes in the white adipose tissue (WAT), called recruitment, might play an important role in the prevention of obesity. The recruitment phenomenon has until now been investigated mostly in vivo. OBJECTIVES: This study is an attempt to mimic in vitro the recruitment phenomenon. It consisted in culturing the stroma vascular fractions of mouse BAT and WAT in a brown adipocyte differentiation medium. The multilocular cells obtained, referred to as BAT(B) and WAT(B) adipocytes, respectively, were compared. RESULTS: The BAT(B) and WAT(B) adipocytes were morphologically different. The expressions of UCP1, peroxisome proliferator-activated receptor-gamma coactivator-1alpha (PGC-1alpha), leptin and resistin mRNAs were low in WAT(B) adipocytes as compared with those in BAT(B) adipocytes. The expressions of UCP1 and PGC-1alpha proteins were, however, much higher in WAT(B) adipocytes, amounting 51% and 36% of those in BAT(B) adipocytes. The patterns of expression of UCP1, PGC-1alpha and leptin in the BAT(B) and in WAT(B) adipocytes were different with a higher relative expression of PGC-1alpha in the latter. Rosiglitazone increased UCP1 mRNA expression 4.5-fold in the BAT(B) and significantly more, 7.9-fold, in the WAT(B) adipocytes. Retinoic acid and triiodothyronine increased UCP1 mRNA expression in the BAT(B) adipocytes 1.6- and 2-fold, respectively but, surprisingly, slightly decreased UCP1 mRNA expression in the WAT(B) adipocytes. CONCLUSIONS: The study suggests that the nature and possibly the origin of WAT brown adipocytes is different from that of BAT brown adipocytes. It proposes an in vitro approach that could prove very useful to better characterize the WAT brown adipocyte-like cells.

Effects of leptin on energy metabolism in beta-less mice.

OBJECTIVE: To investigate the impact of beta-adrenoceptor deficiency on the metabolic effects of leptin. MEASUREMENTS: Leptin was infused subcutaneously through an osmotic minipump in wild-type (WT) and beta(1)/beta(2)/beta(3)-adrenoceptor knockout (beta-less) mice and its effects on food intake, energy expenditure, carbohydrate and lipid utilization as well as on the levels of expression of the brown adipose tissue (BAT), thermogenic marker uncoupling protein-1 (UCP1) and type II deiodinase (D2) mRNAs were compared. RESULTS: Leptin treatment decreased food intake by 23% in both the WT and the beta-less mice. In pair-fed animals being used as controls, leptin treatment was found to increase energy expenditure in WT, but not in beta-less mice. No difference was observed in carbohydrate or fat utilization between leptin-treated WT and beta-less mice. Leptin increased UCP1 and D2 mRNA levels in WT mouse BAT 1.7- and 3-fold, respectively, but had no effect on the expression of these genes in beta-less mouse BAT. CONCLUSION: The stimulatory effects of leptin on oxygen consumption, BAT UCP1 and D2 expression require functional beta-adrenoceptors, but its inhibitory effect on food intake and its stimulatory effect on fat utilization is independent of the beta-adrenoceptor signalling.

Ultrafast dynamics in thiophene investigated by femtosecond pump probe photoelectron spectroscopy and theory.

A hybrid of a time-of-flight mass spectrometer and a time-of-flight "magnetic-bottle type" photoelectron (PE) spectrometer is used for fs pump-probe investigations of the excited state dynamics of thiophene. A resonant two-photon ionization spectrum of the onset of the excited states has been recorded with a tunable UV laser of 190 fs pulse width. With the pump laser set to the first intense transition we find by UV probe ionization first a small time shift of the maxima in the PE spectrum and then a fast decay to a low constant intensity level. The fitted time constants are 80+/-10 fs, and 25+/-10 fs, respectively. Theoretical calculations show that upon geometry relaxation the electronic state order changes and conical intersections between excited states exist. We use the vertical state order S1, S2, S3 to define the terms S1, S2, and S3 for the characterization of the electron configuration of these states. On the basis of our theoretical result we discuss the electronic state order in the UV spectra and identify in the photoelectron spectrum the origin of the first cation excited state D1. The fast excited state dynamics agrees best with a vibrational dynamics in the photo-excited S1 (80+/-10 fs) and an ultrafast decay via a conical intersection, presumably a ring opening to the S3 state (25+/-10 fs). The subsequently observed weak constant signal is taken as an indication, that in the gas phase the ring-closure to S0 is slower than 50 ps. An ultrafast equilibrium between S1 and S2 before ring opening is not supported by our data.

Femtosecond dynamics after ionization: 2-phenylethyl-N,N-dimethylamine as a model system for nonresonant downhill charge transfer in peptides.

The cation of 2-phenylethyl-N,N-dimethylamine (PENNA) offers two local sites for the charge: the amine group and 0.7 eV higher in energy the phenyl chromophore. In this paper, we investigate the dynamics of the charge transfer (CT) from the phenyl to the amine site. We present a femtosecond resonant two-color photoionization spectrum which shows that the femtosecond pump laser pulse is resonant in the phenyl chromophore. As shown previously with resonant wavelengths the aromatic phenyl chromophore can be then selectively ionized. Because the state "charge in the phenyl chromophore" is the first excited state in the PENNA cation, it can relax to the lower-energetic state "charge in the amine site". To follow this CT dynamics, femtosecond probe photoabsorption of green light (vis) is used. The vis light is absorbed by the charged phenyl chromophore, but not by the neutral phenyl and the neutral or cationic amine group. Thus, the absorption of vis photons of the probe laser pulse is switched off by the CT process. For detection of the resonant absorption of two or more vis photons in the cation the intensity of a fragmentation channel is monitored which opens only at high internal energy. The CT dynamics in PENNA cations has a time constant of 80 +/- 28 fs and is therefore not a purely electronic process. Because of its structural similarity to phenylalanine, PENNA is a model system for a downhill charge transfer in peptide cations.

Nitric oxide end products in patients hospitalized for diarrhoea.

OBJECTIVE: Increased nitric oxide (NO) has been demonstrated in inflammatory bowel diseases (IBD). Plasma and urinary nitrite and nitrate are usually considered to reflect global NO generation. Recently it has been suggested that plasma nitrate may be a discriminant indicator between infectious enterocolitis (IC) and IBD. To investigate this hypothesis we compared plasma and 24 h urinary nitrite and nitrate in 13 healthy controls, 44 patients with IBD [Crohn's disease (CD) n = 30; ulcerative colitis (UC) n = 14], 16 patients presenting with IC and seven chronic radiation enterocolitis (RE) patients. RESULTS: Despite a trend towards higher plasma nitrate in IC (54.6+/-11.4 micromol/l) than in the other groups (CD: 38.4+/-4.8, UC: 34.8+/-8.4, RE: 34.7+/-7.5, controls: 31.1+/-5.2), this difference was not statistically significant. Urinary nitrate was higher in IBD, IC and RE than in controls, with no difference between these groups. Nitrite concentrations were not different. Nitrate levels were positively correlated with blood and 24 h urinary neopterin (e.g. plasma nitrate and blood neopterin: r = 0.54, P<0.0001), and in some cases, to C-reactive protein. CONCLUSIONS: High nitrate (in our case only urinary nitrate) appears to be secondary to the magnitude of the inflammation rather than the aetiology of the diarrhoea. It should therefore more likely be considered as a marker of the severity of the inflammatory response rather than used as a discriminant indicator between IC and IBD patients.

Electron solvation in finite systems: femtosecond dynamics of iodide. (Water)n anion clusters

Electron solvation dynamics in photoexcited anion clusters of I-(D2O)n=4-6 and I-(H2O)4-6 were probed by using femtosecond photoelectron spectroscopy (FPES). An ultrafast pump pulse excited the anion to the cluster analog of the charge-transfer-to-solvent state seen for I- in aqueous solution. Evolution of this state was monitored by time-resolved photoelectron spectroscopy using an ultrafast probe pulse. The excited n = 4 clusters showed simple population decay, but in the n = 5 and 6 clusters the solvent molecules rearranged to stabilize and localize the excess electron, showing characteristics associated with electron solvation dynamics in bulk water. Comparison of the FPES of I-(D2O)n with I-(H2O)n indicates more rapid solvation in the H2O clusters.

Cortical visual processing is temporally dispersed by luminance in human subjects.

Increasing the intensity of a stimulus such as luminance results in faster processing of the signal and therefore decreases simple motor reaction time (RT). We studied the latencies of visual evoked potentials (VEPs, N80, P100, N130) and RTs in eight subjects to flashing spots of light while varying the luminance of the spots from 1 to 1000 cd/m2. The data show that processing time as a function of intensity is modified not only at the retina but also at later processing sites. This indicates a temporal dispersion of the visual signal over the whole processing stream from visual input all the way to motor output.

[Soluble tumor necrosis factor receptors in patients with systemic lupus erythematosus]. / Récepteurs solubles au tumor necrosis factor chez des malades ayant un lupus systémique.

OBJECTIVE: To study the variations of type II soluble receptors for tumor necrosis factor (sR-TNF) in patients with systemic lupus erythematosus and investigate their use in the clinical setting. PATIENTS AND METHODS: Twenty-six patients with systemic lupus were followed for a mean 3 years. sR-TNF and other immunological parameters (C reactive protein, anti-DNA antibodies, C3 and C4 complement fractions, soluble receptors for interleukin 2) were measured in sera at different points of the disease course. The systemic lupus activity measure (SLAM) was determined at each point, and confronted with the biology results. The study was cross sectional for the group and longitudinal for the patients. RESULTS: sR-TNF was the immunological parameter which correlated best with SLAM. It also correlated with sedimentation rate, C reactive protein, thrombopenia, anemia, creatinine level, anti-DNA antibodies and sR-IL2. The longitudinal study pointed out however that this finding is not consistent for each patient. CONCLUSION: A rise in sR-TNF related to systemic lupus activity but is of limited practical interest for individual patient follow-up.

[Concept and realization of nationally applicable clinic guidelines in the intra- and internet--a quality assurance measure in the health care system?]. / Konzeption und Realisierung überregional nutzbarer Klinikrichtlinien im Intra- und im Internet--eine Qualitätssicherungsmassnahme im Gesundheitssystem?

This project aimed at the realisation of a database for clinical guidelines in abdominal surgery and oncology, which can be used within a hospital network as well as from the internet. The "Klinikmanual Chirurgie" (currently in German only) can be accessed via http://nt1.chir.med.tu-muenchen.de/manual.h tm worldwide and free of charge. An English version is in preparation.

[Ambulatory surgery--definition, socioeconomic and legal aspects]. / Ambulante Chirurgie--Definitionen, sozio-ökonomische und juristische Aspekte.

One of the most important intentions of GSG 1992 is the favourization of traditional ambulatory surgery by legislative measures. However in general surgery a much higher potential for a significant reduction in cases of classical hospitalization and thus concomitant substantial financial savings lies in short-stay eg. one-day/one-night procedures. At present methods of laparoscopic cholecystectomy, thyroid surgery and herniorrhaphy-together about 50% of common hospital patients-are developed enough to be practiced safely under this strategy. In the future further suitable techniques could evolve from minor access surgical concepts.

[Preoperative autologous blood donation and disposition. Possibilities and prerequisites]. / Präoperative Eigenblutspende und-bereitstellung. Möglichkeiten und Voraussetzungen.

Preoperative blood deposit can help to avoid the risks of homologous blood transfusions. However, it is necessary that prerequisites concerning medicine standard, organisation, technical questions and administrative aspects have to be fulfilled.

[Ambulatory surgery at the surgical clinic and polyclinic of the Munich Technical University]. / Tageschirurgie an der chirurgischen Klinik und Poliklinik der Technischen Universität München.

Day-care and especially short-stay surgery were found attractive logistics for surgery of the hand, inguinal hernia repair, laparoscopic cholecystectomy, invasive measures for tumor staging such as diagnostic laparoscopy and implantation of vascular ports for chemotherapy. This new way of surgical organization is a practical alternative to overcome increasing limitations of hospitalization capacities and to conserve everyday surgery which is necessary for the teaching of students and surgical beginners within otherwise highly specialized institutions.

[Technique of laparoscopic cholecystectomy]. / Technik der laparoskopischen Cholezystektomie.

A new technique for laparoscopic cholecystectomy is described which regards as far as possible the conventional approach to cholecystectomy. Positioning of the operating team is identical to normal cholecystectomy, and the surgeon is able to use both hands to operate since the camera is handled by the assistant. Dissection of the infundibulum is performed in the socalled "tease-and-tear" technique. The peritoneal layer is opened by coagulation. The cystic duct and artery are bluntly dissected by a commercially available dissector. Fat and connective tissue are gently torn off from both structures. After closure of cystic duct and artery by clips, the gallbladder is cut out by thermocoagulation. 178 operations were performed by 8 surgeons; mean duration of the operation was 60 min. A change to open cholecystectomy was necessary in 2.5%. Three complications occurred, requiring reoperation in one case of insufficiency of the cystic duct and another one with intestinal perforation. In the third case, bile leakage from an aberrant bile-duct occurred but dried up spontaneously after a few days. No death occurred nor were there any lesions of the common bile-duct. Conclusively, this new technique seems to be safe and simple to teach due to its approximity to the conventional technique and is recommended as a standard procedure.