Versatile optical laser system for experiments at the European X-ray free-electron laser facility.

We present the main features of the final prototype of a pulsed optical laser, developed for pump-probe and other experiments in conjunction with the femtosecond x-ray beams at the European X-ray free-electron laser facility. Adapted to the temporal x-ray emission pattern of the facility, the laser provides 10 Hz bursts of up to 600 µs duration with intra-burst pulse frequencies as high as 4.5 MHz. In this mode, we have generated pulses as short as 12 fs at 350 W average power during the burst and with beam qualities close to the diffraction limit. This is, to the best of our knowledge, the highest power to date of a few-cycle laser operating at a center wavelength of 800 nm. Important for experimental flexibility, the laser can be configured in various unique ways, enabling, e.g., energy scaling to >3 mJ per pulse through a frequency change down to 100 kHz and the generation of nearly transform limited pulses between 12 fs and 300 fs. In addition to the 800 nm femtosecond beam line, a synchronized long pulse (0.8 ps or 400 ps) 1030 nm beam can be utilized, offering up to 4 kW burst average power, i.e. up to 40 mJ per pulse at 100 kHz. Efficient nonlinear wavelength conversion and tuning through intrinsic and external means further enhance the capabilities of the laser.

High power burst-mode optical parametric amplifier with arbitrary pulse selection.

We present results from a unique burst-mode femtosecond non-collinear optical parametric amplifier (NOPA) under development for the optical - x-ray pump-probe experiments at the European X-Ray Free-Electron Laser Facility. The NOPA operates at a burst rate of 10 Hz, a duty cycle of 2.5% and an intra-burst repetition rate of up to 4.5 MHz, producing high fidelity 15 fs pulses at a center wavelength of 810 nm. Using dispersive amplification filtering of the super-continuum seed pulses allows for selectable pulse duration up to 75 fs, combined with a tuning range in excess of 100 nm whilst remaining nearly transform limited. At an intra-burst rate of 188 kHz the single pulse energy from two sequential NOPA stages reached 180 µJ, corresponding to an average power of 34W during the burst. Acousto- and electro-optic switching techniques enable the generation of transient free bursts of required length and the selection of arbitrary pulse sequences inside the burst.

Compact picosecond mode-locked and cavity-dumped Nd:YVO4 laser.

We report on a diode pumped, semiconductor saturable absorber mirror mode-locked picosecond Nd:YVO(4) oscillator with cavity-dumping. In pure cw-mode-locking this laser produced up to 17W of average power at a pulse repetition rate of 9.7MHz, corresponding to a pulse energy of 1.7microJ. Using an electro-optic cavity dumper, we achieved average powers up to 7.8W at 500kHz and 10W at 1MHz dumping rate. With corresponding pulse energies of 15.6microJ and 10microJ respectively and pulsewidths around 10ps, this laser could become a compact source for materials processing applications, alternative to more complex schemes such as regenerative amplifiers or ultra-long resonator oscillators.

High power cw and mode-locked oscillators based on Yb:KYW multi-crystal resonators.

We report on a high power diode-pumped laser using multiple bulk Yb:KY(WO(4))(2) (KYW) crystals in a resonator optimised for this operation. From a dual-crystal resonator we obtain more than 24 W of cw-power in a TEM(00) mode limited by the available pump power. We also present results for semiconductor saturable absorber mirror (SESAM) mode-locking in the soliton as well as positive dispersion regime with average output powers of 14.6 W and 17 W respectively.

Expression of the RNA-binding protein IMP1 correlates with poor prognosis in ovarian carcinoma.

The IMP (IGFII mRNA-binding protein) family comprises a group of three RNA-binding proteins involved in the regulation of cytoplasmic mRNA-fate. Recent studies identified IMP proteins as oncofetal factors in various neoplasias, but knowledge of a potential role in ovarian carcinomas is still lacking. The immunohistochemical analysis of 107 ovarian carcinomas, 30 serous borderline tumors of the ovary and five normal ovaries revealed de novo synthesis of IMP1 in 69% of ovarian carcinomas. Elevated IMP1 expression was observed preferentially in high-grade and high-stage cases and was a significant prognostic indicator for reduced recurrence-free and overall survival. Phenotypic studies in ovarian carcinoma-derived ES-2 cells demonstrated that IMP1 knockdown affects proliferation and cell survival. Reduced proliferation was associated with decreased c-myc mRNA half-life, suggesting IMP1 as an oncogenic factor that is involved in promoting elevated proliferation by stabilizing the c-myc mRNA in ovarian carcinoma cells.

Association between tPA therapy and raised early matrix metalloproteinase-9 in acute stroke.

BACKGROUND: Matrix metalloproteinase-9 (MMP9) is expressed in acute ischemic stroke and up-regulated by tissue plasminogen activator (tPA) in animal models. The authors investigated plasma MMP9 and its endogenous inhibitor, tissue inhibitor of metalloproteinase (TIMP1), in tPA-treated and -untreated stroke patients. METHODS: Nonstroke control subjects and consecutive ischemic stroke patients presenting within 8 hours of onset were enrolled. Blood was sampled within 8 hours and at 24 hours, 2 to 5 days and 4 to 6 weeks. MMP9 and TIMP1 were analyzed by ELISA and gel zymography. RESULTS: Fifty-two cases (26 tPA treated, 26 tPA untreated) and 27 nonstroke control subjects were enrolled. Hyperacute MMP9 was elevated in tPA-treated vs tPA-untreated patients (medians 43 vs 28 ng/mL; p = 0.01). tPA therapy independently predicted hyperacute MMP9 after adjustment for stroke severity, volume, and hemorrhagic transformation (p = 0.01). There was a trend toward lower hyperacute TIMP1 levels in tPA-treated vs tPA-untreated patients (p = 0.06). Hyperacute MMP9 was correlated to poor 3-month modified Rankin Scale outcome (r = 0.58, p = 0.0005). CONCLUSION: Tissue plasminogen activator independently predicted plasma matrix metalloproteinase-9 (MMP9) in the first 8 hours after human ischemic stroke. As MMP9 may be an important mediator of hemorrhagic transformation, alternative thrombolytic agents or therapeutic MMP9 inhibition may increase the safety profile of acute stroke thrombolysis.

Supercontinuum generation system for optical coherence tomography based on tapered photonic crystal fibre.

We report smooth and broad continuum generation using a compact femtosecond Ti:Sapphire laser as a pump source and a tapered photonic crystal fibre as a nonlinear element. Spectral output is optimized for use in optical coherence tomography, providing a maximum longitudinal resolution of 1.5 microm in free space at 809 nm centre wavelength without use of additional spectral filtering.

C-band waveguide amplifier produced by femtosecond laser writing.

An optical waveguide amplifier fabricated on erbium-ytterbium-doped phosphate glass by direct femtosecond laser writing is demonstrated. The waveguides are manufactured using 1040-nm radiation from a diode-pumped cavity-dumped Yb:KYW oscillator, operating at a 885 kHz repetition rate, with a 350 fs pulse duration. Peak internal gain of 9.2 dB is obtained at 1535 nm, with a minimum internal gain of 5.2 dB at 1565 nm. Relatively low insertion losses of 1.9 dB enable for the first time an appreciable net gain in the full C-band of optical communications.

Diode-pumped femtosecond laser oscillator with cavity dumping.

We report on a diode-pumped tunable Yb:glass femtosecond laser oscillator with electro-optic cavity dumping. Pulses with energies exceeding 400 nJ and peak powers greater than 1 MW were generated at repetition frequencies as high as 200 kHz. This laser forms a compact light source for various scientific and industrial applications such as micromachining.

Passive mode locking of a Nd:YVO4 laser with an extra-long optical resonator.

We describe a mode-locked, diode-pumped Nd:YVO4 laser with a very long optical cavity operating at 1064 nm. High-modulation, InGaAs quantum-well, semiconductor saturable-absorber mirrors were used for passive mode locking, providing a stable train of 13-ps pulses. A novel zero-q-transformation multipass cell provided a variable-length optical cavity as much as 100 m long. The output beam had M2 < 1.1 at average powers of 4.1, 3.9, and 3.5 W at repetition rates of 4.1, 2.6, and 1.5 MHz, respectively. To the best of our knowledge the last of these is the lowest repetition rate ever generated directly from a mode-locked nonfiber solid-state laser without cavity dumping.

Passive mode locking of a self-frequency-doubling Yb:YAl(3) (BO(3))(4) laser.

We report passive mode-locking experiments with a novel self-doubling laser crystal Yb:YAl(3)(BO(3))(4) (Yb:YAB). The diode-pumped laser was mode locked by an ion-implanted semiconductor saturable absorber mirror. Far off phase matching, soliton mode locking produced pulse widths of 198 fs to 1.4 ps, with up to 660-mW output and optical efficiency of 24% at 1040 nm. The shortest pulses had a peak power of 28 kW with 440-mW average power and 16% efficiency. A few degrees off phase matching, a total of 60 mW of green femtosecond pulses was generated simultaneously. Close to phase matching, the laser produced picosecond pulses and, without infrared output, a total of 270 mW of green output, corresponding to 10% conversion efficiency (absorbed pump to green output).

Generation of sub-10-fs pulses from a Kerr-lens mode-locked Cr(3+):LiCAF laser oscillator by use of third-order dispersion-compensating double-chirped mirrors.

Pulses as short as 9 fs at 220-mW average power and a 97-MHz repetition rate are generated from a cw Ti:sapphire-pumped Kerr-lens mode-locked Cr(3+)LiCAF laser oscillator employing broadband double-chirped mirrors for second- and third-order dispersion compensation. Fine adjustment of dispersion is accomplished with a fused-silica prism pair. The result demonstrates that Raman-induced self-frequency shifting of the pulse does not limit sub-10-fs pulse generation from colquiriite crystals.

All-optical active mode locking with a nonlinear semiconductor modulator.

All-optical active mode locking of a picosecond Nd:YVO(4) laser is demonstrated by use of an intracavity semiconductor nonlinear Fabry-Perot mirror. The reflectivity of the Fabry-Perot mirror is modulated by optical carrier injection. Depending on the carrier recombination time, the width of the Nd:YVO(4) laser pulses varies from 6 to 20 ps, as is typical for passively mode-locked Nd:YVO(4) lasers.

[Effect of graduate education on complication rate and costs of hip prosthesis implantation]. / Einfluss der Weiterbildung auf Komplikationsdichte und Kosten einer Hüftendoprothese.

PURPOSE: We studied the effect of surgical experience with regard to complications and costs after 3290 primary total hip arthroplasties (diagnosis: osteoarthritis). METHODS: We retrospectively analysed duration of clinical treatment and anaesthesia, blood loss, number of banked blood and perioperative complications (fractures of the shaft and acetabulum, dislocations, peripheral nerve injuries, thrombosis of the deep veins, embolism and deep wound infections) related to the experience of the surgeon (junior surgeons in comparison to the consultant) after 3290 primary THA's from 1974 to 1997. Cost analysis was performed by measurement of time and material expenditure. RESULTS: 39.2% of all operations were performed by junior surgeons. This number emphasizes the realisation of the order of education in our clinic. Both groups showed just few differences in number of duration of clinical treatment, banked blood, nerve leasions and embolism. We found statistically not significant increased rates of fractures and dislocations in the THA's performed by an unexperienced surgeon, however the rates decreased the last 14 years. Significantly higher rates of deep wound infection at the beginning decreased in the last 9 years to the level of experienced surgeons. Due to longer operation times, we calculated higher expenses of DM 85 per case in the group of junior surgeons. CONCLUSIONS: Using an educational program, carried out with an experienced surgeon, there is nearly no additional risk for the patient regarding the complication rates in THA's. However, there are higher financial expenses that have to be calculated.

Identification and quantification of phosphatidylethanolamine-derived glucosylamines and aminoketoses from human erythrocytes--influence of glycation products on lipid peroxidation.

While the Maillard reaction of amino acids and proteins as well as its consequences in vivo has been thoroughly investigated, little attention has so far been paid to the glycation of aminophospholipids such as phosphatidylethanolamine (PE) or phosphatidylserine (PS), which are essential for structure and functionality of biological membranes. PE-derived glucosylamines (Schiff-PEs) and aminoketoses (Amadori-PEs) have now for the first time been simultaneously identified and quantified in erythrocytes from diabetics and healthy individuals by liquid chromatography-electrospray mass spectrometry (LC-(ESI)MS). The amounts of glycated PE (gPE) were significantly higher in diabetics (0.18-34.2 mol% Schiff-PE and 0.047-0.375 mol% Amadori-PE) than in controls (0.12-3.99 mol% Schiff-PE and 0.018-0.055 mol% Amadori-PE). A positive correlation between fructosylated hemoglobin (HbA(1c)) and the gPE levels was established. No advanced glycation endproducts (AGEs) like 5-hydroxymethylpyrrole-2-carbaldehyde (pyrrole-PE), carboxymethyl (CM-PE), or carboxyethyl (CE-PE) derivatives were detected. To investigate the influence of gPE on lipid peroxidation of biological membranes, liposomes consisting of soy-PE and synthetically prepared Amadori-PE (16:0-16:0) were incubated for several days and the formation of oxidation products was monitored. It could be shown that Amadori-PE extensively promotes lipid peroxidation even in the absence of transition metal ions like Cu(2+) and Fe(3+). Oxidative damage to membrane lipids therefore is supposed to be at least partially caused by the glycation of aminophospholipids.

Identification and quantitative evaluation of the lysine-arginine crosslinks GODIC, MODIC, DODIC, and glucosepan in foods.

The presence of the various protein crosslinks GOLD 2, MOLD 3, GODIC 4, MODIC 5, DODIC 6, and glucosepan 7 in foods has been established for the first time by liquid chromatography-mass spectrometry (LC-MS) with electrospray ionization (ESI). In compounds 2 and 3 two lysine moieties, in 4-7 a lysine and an arginine side chain are joined by the crosslink. Unequivocal identification of 2-7 was achieved with independently synthesized reference material. The quantitative results for the investigated foodstuffs show MODIC 5 to be the most important Maillard crosslink. The concentrations of 5 and GODIC 4 are 5-10 fold higher than those of the corresponding imidazolium compounds 3 and 2, establishing 5 and 4 as the major food protein crosslinks derived from methylglyoxal and glyoxal, respectively. The maximum value of 151 mg MODIC 5/kg protein (equivalent to 0.42 mmol/kg protein) was found in a butter biscuit sample which also shows the highest overall Maillard crosslink content with 0.71 mmol 47/kg protein. These first quantitative results suggest that compounds 4-7 can be jointly responsible for protein polymerization in the course of food processing.

Identification and characterization of a mammalian enzyme catalyzing the asymmetric oxidative cleavage of provitamin A.

In vertebrates, symmetric versus asymmetric cleavage of beta-carotene in the biosynthesis of vitamin A and its derivatives has been controversially discussed. Recently we have been able to identify a cDNA encoding a metazoan beta,beta-carotene-15,15'-dioxygenase from the fruit fly Drosophila melanogaster. This enzyme catalyzes the key step in vitamin A biosynthesis, symmetrically cleaving beta-carotene to give two molecules of retinal. Mutations in the corresponding gene are known to lead to a blind, vitamin A-deficient phenotype. Orthologs of this enzyme have very recently been found also in vertebrates and molecularly characterized. Here we report the identification of a cDNA from mouse encoding a second type of carotene dioxygenase catalyzing exclusively the asymmetric oxidative cleavage of beta-carotene at the 9',10' double bond of beta-carotene and resulting in the formation of beta-apo-10'-carotenal and beta-ionone, a substance known as a floral scent from roses, for example. Besides beta-carotene, lycopene is also oxidatively cleaved by the enzyme. The deduced amino acid sequence shares significant sequence identity with the beta,beta-carotene-15,15'-dioxygenases, and the two enzyme types have several conserved motifs. To establish its occurrence in different vertebrates, we then attempted and succeeded in cloning cDNAs encoding this new type of carotene dioxygenase from human and zebrafish as well. As regards their possible role, the apocarotenals formed by this enzyme may be the precursors for the biosynthesis of retinoic acid or exert unknown physiological effects. Thus, in contrast to Drosophila, in vertebrates both symmetric and asymmetric cleavage pathways exist for carotenes, revealing a greater complexity of carotene metabolism.

Formation pathways for lysine-arginine cross-links derived from hexoses and pentoses by Maillard processes: unraveling the structure of a pentosidine precursor.

Covalently cross-linked proteins are among the major modifications caused by the advanced Maillard reaction. So far, the chemical nature of these aggregates and their formation pathways are largely unknown. Synthesis and unequivocal structural characterization are reported for the lysine-arginine cross-links N(6)-(2-([(4S)-4-ammonio-5-oxido-5-oxopentyl]amino)-5-[(2S,3R)-2,3,4- trihydroxybutyl]-3,5-dihydro-4H-imidazol-4-ylidene)-l-lysinate (DOGDIC 12), N(6)-(2-([(4S)-4-ammonio-5-oxido-5-oxopentyl]amino)-5-[(2S)-2,3-dihydroxypropyl]-3,5-dihydro-4H-imidazol-4-ylidene)-l-lysinate (DOPDIC 13), and 6-((6S)-2-([(4S)-4-ammonio-5-oxido-5-oxopentyl] amino)-6-hydroxy-5,6,7,7a-tetrahydro-4H-imidazo[4,5-b] pyridin-4-yl)-l-norleucinate (pentosinane 10). For these compounds, as well as for glucosepane 9 and pentosidine 11, the formation pathways could be established by starting from native carbohydrates, Amadori products, and 3-deoxyosones, respectively. Pentosinane 10 was unequivocally proven to be an important precursor of pentosidine 11, which is a well established fluorescent indicator for advanced glycation processes in vivo. The Amadori products are shown to be the pivots in the formation of the various cross-links 9-13. The bicyclic structures 9-11 are directly derived from aminoketoses, whereas 12 and 13 stem from reaction with the 3-deoxyosones. All products 9-13 were identified and quantified from incubations of bovine serum albumin with the respective 3-deoxyosone or carbohydrate. From these results it seems fully justified to expect both glucosepane 9 and DOGDIC 12 to constitute important in vivo cross-links.

Generation of 5-fs pulses and octave-spanning spectra directly from a Ti:sapphire laser.

Spectra extending from 600 to 1200 nm have been generated from a Kerr-lens mode-locked Ti:sapphire laser producing 5-fs pulses. Specially designed double-chirped mirror pairs provide broadband controlled dispersion, and a second intracavity focus in a glass plate provides additional spectral broadening. These spectra are to our knowledge the broadest ever generated directly from a laser oscillator.

Formation of a phospholipid-linked pyrrolecarbaldehyde from model reactions of D-glucose and 3-deoxyglucosone with phosphatidyl ethanolamine.

Phospholipid-linked 'advanced glycation end products' (AGEs) are supposed to play an important role for lipid oxidation in vivo. The identification of the pyrrolecarbaldehyde 1-[2-formyl-5-(hydroxymethyl)-1 H-pyrrol-1-yl]-4,10-dioxo-7-(tetradecanoyloxy)-3,5,9-trioxa- 4lambda5-phosphatricosan-4-olate (7) from model reactions of D-glucose or 3-deoxyglucosone (4, 3-DG) with phosphatidyl ethanolamine (PE) is described. A preparation method is given for 1-(2-hydrox¿ethyl)-5-(hydroxymethyl)-1H-pyrrole-2-carbaldehyde (8). Independent syntheses as well as unequivocal structural characterization are reported for the substitution products of 8 1-(2-hydroxyethyl)-5-(methoxymethyl)-1H-pyrrole-2-carbaldehyde (9a) and 5-(ethoxymethyl)-1-(2-hydroxyethyl)-1H-pyrrole-2-carbaldehyde (9b). For all these compounds, chromatographic and spectroscopic data were established by GLC-MS and HPLC with diode array detection (DAD). PE and D-glucose or 3-DG 4 were either incubated at pH 7.4, 100 degrees C for 3 h or at pH 7.4, 37 degrees C for 5 weeks in neat buffer or ethanol buffer mixtures. The phospholipid fraction was purified on a C18 solid-phase extraction column and cleaved with ethanolic potassium hydroxide. The carbaldehyde 8, released in this process, was identified bs GLC-MS and quantified by HPLC-DAD. Formation of 7 is favored in the ethanol buffer reactions relative to those in buffer solution only although the amounts determined from the 37 degrees C incubations generally are very low. It seems likely, therefore, that phospholipid-linked pyrrolecarbaldehydes, such as 7, are biomarkers rather than effectors of membrane damage in vivo.