Biomedical subjects

Frank Müller

Publications and source records attributed to Frank Müller.

At least 19 recordsLinked to original sources

PubMed2006

The shear modulus of the human vocal fold, preliminary results from 20 larynxes.

Quantification of the elastic properties of the human vocal fold provides invaluable data for researchers deriving mathematical models of phonation, developing tissue engineering therapies, and as normative data for comparison between healthy and scarred tissue. This study measured the shear modulus of excised cadaver vocal folds from 20 subjects. Twenty freshly excised human larynxes were evaluated less than four days post-mortem. They were split along the saggital plane and mounted without tension. Shear modulus was obtained by two different methods. For method 1 cyclical shear stress was applied transversely to the mid-membranous portion of the vocal fold, and shear modulus derived by applying a simple shear model. For method 2 the apparatus was configured as an indentometer, and shear modulus obtained from the stress/strain data by applying an established analytical technique. Method 1 shear model for male larynxes yielded a range from 246 to 3,356 Pa, with a mean value of 1,008 and SD of 380. The range for female larynxes was 286-3,332 Pa, with a mean value of 1,237 and SD of 768. Method 2 indentometer model for male larynxes yielded a range from 552 to 2,741 Pa, with a mean value of 1,000 and SD of 460. The range for female larynxes was 509-1,989 Pa, with a mean value of 1,332 and SD of 428. We have successfully demonstrated two methodologies that are capable of directly measuring the shear modulus of the human vocal fold, without dissecting out the vocal fold cover tissue. The sample size of nine female and 11 male larynxes is too small to validate a general conclusion. The high degree of variability in this small cohort of subjects indicates that factors such as age, health status, and post-mortem delay may be significant; and that there is range of 'normality' for vocal fold tissue.

Biomechanical Phenomena↗

PubMed2006

Quantum dots--a versatile tool in plant science?

An optically stable, novel class of fluorophores (quantum dots) for in situ hybridisation analysis was tested to investigate their signal stability and intensity in plant chromosome analyses. Detection of hybridisation sites in situ was based on fluorescence from streptavidin-linked inorganic crystals of cadmium selenide. Comparison of quantum dots (QDs) with conventional detection systems (Alexa 488) in immunolabeling experiments demonstrated greater sensitivity than the conventional system. In contrast, detection of QDs in in situ hybridisation of several plant chromosomes, using several high-copy sequences, was less sensitive than Alexa 488. Thus, semiconductor nanocrystal fluorophores are more suitable for immunostaining but not for in situ hybridisation of plant chromosomes.

Journal Article↗

PubMed2006

Hydroxyapatite growth on anodic TiO2 nanotubes.

In the present work, we study the growth of hydroxyapatite formation on different TiO(2) nanotube layers. The nanotube layers were fabricated by electrochemical anodization of titanium in fluoride-containing electrolytes. To study various nanotube lengths, layers with an individual tube diameter of 100 nm were grown to a thickness of approximately 2 mum or 500 nm. The ability to form apatite on the nanotube layers was examined by immersion tests combined with SEM, XRD and FT-IR investigations. For reference, experiments were also carried out on compact anodic TiO(2) layers. The results clearly show that the presence of the nanotubes on a titanium surface enhances the apatite formation and that the 2-mum thick nanotube layer triggers deposition faster than the thinner layers. Tubes annealed to anatase, or a mixture of anatase and rutile are clearly more efficient in promoting apatite formation than the tubes in their "as-formed" amorphous state.

Coated Materials, Biocompatible↗

PubMed2006

The bHLH transcription factor hand2 is essential for noradrenergic differentiation of sympathetic neurons.

The basic helix-loop-helix transcription factor Hand2, together with Ascl1, Phox2a, Phox2b and Gata2/Gata3, is induced by bone morphogenetic proteins in neural crest-derived precursor cells during sympathetic neuron generation. Hand2 overexpression experiments and the analysis of its function at the Dbh promotor implicated Hand2 in the control of noradrenergic gene expression. Using the zebrafish hand2 deletion mutant hands off, we have now investigated the physiological role of hand2 in the development of sympathetic ganglia. In hands off mutant embryos, sympathetic precursor cells aggregate to form normal sympathetic ganglion primordia characterized by the expression of phox2b, phox2a and the achaete-scute family member zash1a/ascl1. The expression of the noradrenergic marker genes th and dbh is strongly reduced, as well as the transcription factors gata2 and tfap2a (Ap-2alpha). By contrast, generic neuronal differentiation seems to be unaffected, as the expression of elavl3 (HuC) is not reduced in hands off sympathetic ganglia. These results demonstrate in vivo an essential and selective function of hand2 for the noradrenergic differentiation of sympathetic neurons, and implicates tfap2a and gata2 as downstream effectors.

Animals↗

PubMed2005

Offspring from non-stimulated calves at an age younger than two months: a preliminary report.

This study investigated the viability of embryos from non-stimulated 2-3-month-old calves generated in vitro using oocytes from follicles of defined size in terms of their ability to produce full-term pregnancies. Ablation of follicles>or=4 mm was used to induce the emergence of a new follicular wave, and calves (n=3) were laparoscopically punctured three times at 7-day-intervals to recover cumulus-oocyte-complexes (COCs) from follicles>8 (group A) and between 4-8 mm (group B). Calves were aged 49, 56, and 80 days, respectively, at first recovery. Morphologically intact COCs were subjected to in vitro maturation, fertilization, and embryo culture, and compact morulae/blastocysts were transferred on day 7 post-insemination to synchronized virgin heifers. Blood typing was used for maternity analysis. A total of 29 COCs were recovered, 21 cultured, yielding 11 cleaved embryos (52.4%) and 6 compact morulae/blastocysts (28.6%). No differences were observed between groups. Transfer of the 6 embryos to 5 recipients resulted in three pregnancies (one from group A and two from group B). Two normal male offspring (both from group B), with birth weights of 44 and 51 kg, were born, and two donor calves, aged 56 and 59 days, were identified as the dams. In conclusion, the results demonstrate that embryos generated in vitro from oocytes from non-stimulated calves at an age younger than two months are viable in terms of their ability to produce full-term pregnancies, and suggest that the viability of calf embryos is not related to follicle size.

Animals↗

PubMed2005

Cardiolipin synthase of Arabidopsis thaliana.

Functional expression studies in microorganisms showed that the Arabidopsis thaliana gene At4g04870 represents the cardiolipin synthase (CLS) gene encoding a hydrophobic preprotein of 38 kDa with a cleavable signal peptide for the import into mitochondria. CLS of Arabidopsis over-expressed in Escherichia coli has an alkaline pH optimum, a strict requirement for divalent cations and a distinctly lower K(m) for cytidinediphosphate-diacylglycerol than for phosphatidylglycerol. It displayed a preference for both its substrates esterified with unsaturated acyl groups. Solubilization and purification experiments revealed that the protein requires a defined phospholipid environment, particularly the presence of cardiolipin, to acquire its catalytically active conformation.

Arabidopsis↗

PubMed2005

Ipangulines and minalobines, chemotaxonomic markers of the infrageneric Ipomoea taxon subgenus Quamoclit, section Mina.

A comprehensive GC-MS analysis of 8 Ipomoea species belonging to the subgenus Quamoclit, section Mina revealed that the members of this taxon form combinations of two necine bases with rare necic acids resulting in unique pyrrolizidine alkaloids. The occurrence and diversity of these metabolites show remarkable variations: Some species, especially Ipomoea hederifolia and Ipomoea lobata, are able to synthesize a large number of alkaloids whereas others, especially Ipomoea coccinea and Ipomoea quamoclit, are poor synthesizers with only a few compounds. However, these metabolites are apparently chemotaxonomic markers of this infrageneric taxon in general. They represent either esters of (-)-platynecine (altogether 48 ipangulines and 4 further esters including results of a previous study) or esters of (-)-trachelanthamidine, an additional novel structural type called minalobines (altogether 21 alkaloids). Both types are characterized by section-specific rare necic acids, e.g., ipangulinic/isoipangulinic acid, phenylacetic acid. The alkaloids of Ipomoea cholulensis, I. coccinea, I. hederifolia, Ipomoea neei, and Ipomoea quamoclit were mono and diesters of platynecine. Minalobines turned out to be metabolites of I. lobata (Cerv.) Thell. (syn.: Mina lobata Cerv.) lacking ipangulines. The major alkaloid of this species, minalobine R, has been isolated and identified as 9-O-(threo-2-hydroxy-2-methyl-3-phenylacetoxy-butyryl)-(-)-trachelanthamidine on the basis of spectral data. Apparently only two of the species included in this study, Ipomoea cristulata and Ipomoea sloteri, are able to synthesize both, ipangulines as well as minalobines. Minalobine O could be isolated as a major alkaloid of I. cristulata, its structure has been established as 9-O-(erythro-2-hydroxy-2-methyl-3-tigloyloxy-butyryl)-(-)-trachelanthamidine on the basis of spectral data.

Gas Chromatography-Mass Spectrometry↗

PubMed2005

Chemotaxonomy of the pantropical genus Merremia (Convolvulaceae) based on the distribution of tropane alkaloids.

The occurrence and distribution of tropane and biogenetically related pyrrolidine alkaloids in 18 Merremia species of paleo-, neo-, and pantropical occurrence have been studied. The extensive GC-MS study included members of almost all sections of the genus and has been carried out with epigeal vegetative parts as well as with roots. It comprises altogether 74 tropanes and 13 pyrrolidines including nicotine. Along with datumetine known already from a solanaceous species, the study led to the isolation (from M. dissecta and M. guerichii, respectively) and structure elucidation (spectral data) of four novel 3alpha-acyloxytropanes, merresectines A-D: 3alpha-(4-methoxybenzoyloxy)nortropane (A), 3alpha-kurameroyloxytropane (B), 3alpha-nervogenoyloxytropane (C), 3alpha-[4-(beta-D-glucopyranosyloxy)-3-methoxy-5-(3-methyl-2-butenyl)benzoyloxy]tropane (beta-d-glucoside of D). Moreover, the novel 3alpha,6beta-di-(4-methoxybenzoyloxy)tropane (merredissine) has been isolated from M. dissecta and structurally elucidated. In addition the structures of datumetine and merresectine A could be confirmed by synthesis. Spectral data for two known 3alpha-acyloxytropanes (merresectine E beta-D-glucoside, 4'-dihydroconsabatine) and one known 3beta-acyloxytropane (concneorine) are documented for the first time. The structures of three further merresectines (F-H) have been determined by mass spectrometry. Furthermore, the linkage (2',3- and 2',4-, respectively) of two position isomer N-methylpyrrolidinylhygrines was proven by synthesis. The results of the study contribute to the solution of infrageneric taxonomic problems. Whereas all species yield pyrrolidine alkaloids without suitably differentiating results the diverging occurrence of tropane alkaloids leads to three groups of sections: (1) taxa free of tropanes, (2) taxa with simple tropanes, and (3) taxa with merresectines in addition to simple tropanes.

Alkaloids↗

PubMed2004

Can body volume be determined by PET?

PURPOSE: To avoid dependence on body weight, the standardised uptake value (SUV) in positron emission tomography (PET) can instead be normalised to the lean body mass (LBM), which can be determined from body volume and mass. This study was designed to answer the following questions: Firstly, can the total body volume in principle be determined using PET? Secondly, is the precision of this measurement comparable to that achieved using an established standard method. METHODS: Ten patients were examined during oncological whole-body PET examinations. The whole-body volume of the patients was determined from the transmission scan in PET. Air displacement plethysmography with BOD POD was used for comparison as the standard method of volume determination. RESULTS: In all patients, the whole-body volumes could be determined using PET and the standard method. Bland and Altman [23] analysis for agreement between the volumes determined by the two methods (presentation of differences vs means) revealed a very small difference of -0.14 l. With a mean patient volume of 71.81+/-15.93 l, the relative systematic error is only <0.1%. On this basis, equality of the volume values determined by the two methods can be assumed. CONCLUSION: PET can be used as a supplementary method for experimental determination of whole-body volume and total body fat in tumour patients. The fat content can be used to calculate the LBM and to determine body weight-independent SUVs (SUV(LBM)).

Anthropometry↗

PubMed2004

Essential role of Gata transcription factors in sympathetic neuron development.

Sympathetic neurons are specified during their development from neural crest precursors by a network of crossregulatory transcription factors, which includes Mash1, Phox2b, Hand2 and Phox2a. Here, we have studied the function of Gata2 and Gata3 zinc-finger transcription factors in autonomic neuron development. In the chick, Gata2 but not Gata3 is expressed in developing sympathetic precursor cells. Gata2 expression starts after Mash1, Phox2b, Hand2 and Phox2a expression, but before the onset of the noradrenergic marker genes Th and Dbh, and is maintained throughout development. Gata2 expression is affected in the chick embryo by Bmp gain- and loss-of-function experiments, and by overexpression of Phox2b, Phox2a, Hand2 and Mash1. Together with the lack of Gata2/3 expression in Phox2b knockout mice, these results characterize Gata2 as member of the Bmp-induced cluster of transcription factors. Loss-of-function experiments resulted in a strong reduction in the size of the sympathetic chain and in decreased Th expression. Ectopic expression of Gata2 in chick neural crest precursors elicited the generation of neurons with a non-autonomic, Th-negative phenotype. This implies a function for Gata factors in autonomic neuron differentiation, which, however, depends on co-regulators present in the sympathetic lineage. The present data establish Gata2 and Gata3 in the chick and mouse, respectively, as essential members of the transcription factor network controlling sympathetic neuron development.

Animals↗

PubMed2004

Parts per trillion sensitivity for ethane in air with an optical parametric oscillator cavity leak-out spectrometer.

Spectroscopic detection of ethane in the 3-microm wavelength region was performed by means of a cw optical parametric oscillator and cavity leak-out. We achieved a minimum detectable absorption coefficient of 1.6 x 10(-10) cm 1/square root of Hz, corresponding to an ethane detection limit of 6 parts per trillion/square root of Hz. For 3-min integration time the detection limit was 0.5 parts per trillion. The levels are to our knowledge the best demonstrated so far. These frequency-tuning capabilities facilitated multigas analysis with simultaneous monitoring of ethane, methane, and water vapor in human breath.

Journal Article↗

PubMed2004

Near-field visualization of light confinement in a photonic crystal microresonator.

By using scanning near-field optical microscopy, we directly map the subwavelength confinement of light around a point defect in a two-dimensional photonic crystal microresonator. Comparison of our results with the outcome of three-dimensional finite-difference time domain calculations allows us to identify small imperfections in the structure that result in the spatial modification of the intensity distribution.

Journal Article↗

PubMed2004

Different effects of exercise on plasma concentrations of nebivolol, bisoprolol and carvedilol.

BACKGROUND: In-vitro studies have shown that beta-blockers are taken up into and released from adrenergic cells together with epinephrine and norepinephrine. Consequently, studies in humans revealed an increase in plasma concentrations of propranolol and atenolol, whereas those of carvedilol were not affected by physical exercise. However, nebivolol and bisoprolol never were investigated on this issue. METHODS: Ten healthy males received oral doses of 5 mg nebivolol, 5 mg bisoprolol, and 50 mg carvedilol daily for one week in a cross-over fashion. Exercise was performed at 3 hours following oral intake of the respective last drugs on the eighth day. Blood samples were taken at rest, during the last minute of exercise, and after 15 min of recovery. RESULTS: At rest and during exercise, heart rates were as follows: Nebivolol, 57 +/- 7 and 137 +/- 11 beats/min; bisoprolol, 55 +/- 5 and 139 +/- 14 beats/min; carvedilol, 56 +/- 5 and 135 +/- 13 beats/min, with no significant differences between the drugs. Plasma concentrations were as follows: Nebivolol-rest 0.273 +/- 0.029 ng/ml, exercise 0.274 +/- 0.035 ng/ml, recovery 0.272 +/- 0.035 ng/ml (n.s.). Bisoprolol-rest 4.99 +/- 2.73 ng/ml, exercise 6.49 +/- 5.58 ng/ml, recovery 4.90 +/- 3.06 ng/ml ( p < 0.01). Carvedilol-rest 10.3 +/- 9.3 ng/ml, exercise 9.7 +/- 8.2 ng/ml, recovery 6.5 +/- 5.6 ng/ml ( p < 0.05). DISCUSSION: Plasma concentrations of bisoprolol increased during exercise and returned to baseline during recovery, a behaviour which would have been predicted according to present knowledge. However, exercise had no effect on plasma concentrations of nebivolol and carvedilol, a finding that is in contrast to previous results with other beta-blockers such as propranolol and atenolol. We conclude that both nebivolol and carvedilol are not taken up into and released from adrenergic nerves during exercise, a feature that clearly distinguishes these drugs from other beta-blockers so far investigated.

Administration, Oral↗

PubMed2003

Asymmetric pores in a silicon membrane acting as massively parallel brownian ratchets.

The brownian motion of mesoscopic particles is ubiquitous and usually random. But in systems with periodic asymmetric barriers to movement, directed or 'rectified' motion can arise and may even modulate some biological processes. In man-made devices, brownian ratchets and variants based on optical or quantum effects have been exploited to induce directed motion, and the dependence of the amplitude of motion on particle size has led to the size-dependent separation of biomolecules. Here we demonstrate that the one-dimensional pores of a macroporous silicon membrane, etched to exhibit a periodic asymmetric variation in pore diameter, can act as massively parallel and multiply stacked brownian ratchets that are potentially suitable for large-scale particle separations. We show that applying a periodic pressure profile with a mean value of zero to a basin separated by such a membrane induces a periodic flow of water and suspended particles through the pores, resulting in a net motion of the particles from one side of the membrane to the other without moving the liquid itself. We find that the experimentally observed pressure dependence of the particle transport, including an inversion of the transport direction, agrees with calculations of the transport properties in the type of ratchet devices used here.

Journal Article↗

PubMed2003

Peroxisome degradation requires catalytically active sterol glucosyltransferase with a GRAM domain.

Fungal sterol glucosyltransferases, which synthesize sterol glucoside (SG), contain a GRAM domain as well as a pleckstrin homology and a catalytic domain. The GRAM domain is suggested to play a role in membrane traffic and pathogenesis, but its significance in any biological processes has never been experimentally demonstrated. We describe herein that sterol glucosyltransferase (Ugt51/Paz4) is essential for pexophagy (peroxisome degradation), but not for macroautophagy in the methylotrophic yeast Pichia pastoris. By expressing truncated forms of this protein, we determined the individual contributions of each of these domains to pexophagy. During micropexophagy, the glucosyltransferase was associated with a recently identified membrane structure: the micropexophagic apparatus. A single amino acid substitution within the GRAM domain abolished this association as well as micropexophagy. This result shows that GRAM is essential for proper protein association with its target membrane. In contrast, deletion of the catalytic domain did not impair protein localization, but abolished pexophagy, suggesting that SG synthesis is required for this process.

Autophagy↗

PubMed2003

Arabidopsis phosphatidylglycerophosphate synthase 1 is essential for chloroplast differentiation, but is dispensable for mitochondrial function.

Genetic dissection of the lipid bilayer composition provides essential in vivo evidence for the role of individual lipid species in membrane function. To understand the in vivo role of the anionic phospholipid, phosphatidylglycerol, the loss-of-function mutation was identified and characterized in the Arabidopsis thaliana gene coding for phosphatidylglycerophosphate synthase 1, PGP1. This mutation resulted in pigment-deficient plants of the xantha type in which the biogenesis of thylakoid membranes was severely compromised. The PGP1 gene coded for a precursor polypeptide that was targeted in vivo to both plastids and mitochondria. The activity of the plastidial PGP1 isoform was essential for the biosynthesis of phosphatidylglycerol in chloroplasts, whereas the mitochondrial PGP1 isoform was redundant for the accumulation of phosphatidylglycerol and its derivative cardiolipin in plant mitochondrial membranes. Together with findings in cyanobacteria, these data demonstrated that anionic phospholipids play an important, evolutionarily conserved role in the biogenesis and function of the photosynthetic machinery. In addition, mutant analysis suggested that in higher plants, mitochondria, unlike plastids, could import phosphatidylglycerol from the endoplasmic reticulum.

Arabidopsis↗

PubMed2003

HCN channels are expressed differentially in retinal bipolar cells and concentrated at synaptic terminals.

Hyperpolarization-activated and cyclic nucleotide-gated (HCN) channels codetermine the integrative behaviour of neurons and shape their response to synaptic stimulation. We used immunohistochemistry and patch-clamp recording to study the composition and distribution of HCN channels in the rat retina. All four HCN channel isoforms (HCN1-4) are expressed differentially in the retina. In particular, different classes of bipolar cells have a different inventory of HCN channels. We found no evidence for the formation of heterooligomeric HCN channels. HCN channels are densely clustered at synaptic terminals of bipolar cells and photoreceptors. This suggests that HCN channels are involved in the control of transmitter release.

Animals↗

PubMed2002

Strain and process development for the production of human cytokines in Hansenula polymorpha.

The early status of strain development for the production of interleukin (IL)-6, IL-8, IL-10, and interferon (IFN) gamma is described. The general approach to generating such strains was to amplify gene sequences encoding the mature forms of the various cytokines by PCR from commercially available cDNA sources. The design of the amplificates allowed an in-frame fusion to an MFalpha1 leader segment contained in two basic expression vectors, pFPMT121-MFalpha1 and pTPSMT-MFalpha1. The two vectors differ in that one harbors the methanol-inducible FMD promoter and the other the constitutive TPS1 promoter as control elements for heterologous gene expression. The most advanced process development example is that of IFNalpha-2a. Here, the MOX promoter derived from another key gene of methanol metabolism is used for expression control. The successful development of a production process for Hansenula polymorpha-derived IFNalpha-2a is summarized. This was achieved by combining genetic engineering of suitable production strains with improved processing capabilities for the secreted cytokine, and by purification procedures from cultures grown in yeast extract-peptone-glycerol-based media.

Amino Acid Sequence↗