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1.
J Hosp Infect ; 143: 64-75, 2024 Jan.
Article in English | MEDLINE | ID: mdl-37939884

ABSTRACT

BACKGROUND: Bacteria are becoming increasingly resistant to classical antimicrobial agents, so new approaches need to be explored. AIM: To assess the potential of cold atmospheric plasma for the management of meticillin-resistant Staphylococcus aureus (MRSA). METHODS: The 24, 48, and 72 h resistant and susceptible S. aureus biofilms were exposed to 60, 120, and 180 s treatment with plasma. FINDINGS: Increasing the treatment time results in higher cell reduction for both susceptible and resistant strains of S. aureus (P < 0.05). Up to log10 reduction factor of 5.24 cfu/cm2 can be achieved in 180 s of plasma treatment. Furthermore, plasma can substantially alter the cell's metabolisms and impact cell membrane integrity. However, it has not been shown that plasma can reduce biofilm biomass in the case of 24 h and 48 h biofilms, although the 72 h biofilm was more susceptible, and its biomass was decreased (P < 0.05). The accumulation of intrabacterial reactive oxygen species was also observed, which confirms the plasma's induction of oxidative stress. Finally, it was shown that continuous plasma exposure of bacterial cells does not cause resistance to plasma, nor is resistance developed to cefoxitin. CONCLUSION: Cold atmospheric plasma is a good candidate for S. aureus and MRSA biofilm treatment and may therefore be of value in the bacterial resistance crisis.


Subject(s)
Methicillin-Resistant Staphylococcus aureus , Plasma Gases , Humans , Staphylococcus aureus , Methicillin , Plasma Gases/pharmacology , Disinfection/methods , Biofilms , Anti-Bacterial Agents/pharmacology , Microbial Sensitivity Tests
2.
J Chem Phys ; 153(22): 224104, 2020 Dec 14.
Article in English | MEDLINE | ID: mdl-33317307

ABSTRACT

Photo-induced relaxation processes leading to excimer formations or other traps are in the focus of many investigations of optoelectronic materials because they severely affect the efficiencies of corresponding devices. Such relaxation effects comprise inter-monomer distortions in which the orientations of the monomer change with respect to each other, whereas intra-monomer distortions are variations in the geometry of single monomers. Such distortions are generally neglected in quantum chemical investigations of organic dye aggregates due to the accompanied high computational costs. In the present study, we investigate their relevance using perylene-bisimide dimers and diindenoperylene tetramers as model systems. Our calculations underline the importance of intra-monomer distortions on the shape of the potential energy surfaces as a function of the coupling between the monomers. The latter is shown to depend strongly on the electronic state under consideration. In particular, it differs between the first and second excited state of the aggregate. Additionally, the magnitude of the geometrical relaxation decreases if the exciton is delocalized over an increasing number of monomers. For the interpretation of the vibronic coupling model, pseudo-Jahn-Teller or Marcus theory can be employed. In the first part of this paper, we establish the accuracy of density functional theory-based approaches for the prediction of vibrationally resolved absorption spectra of organic semiconductors. These investigations underline the accuracy of those approaches although shortcomings become obvious as well. These calculations also indicate the strength of intra-monomer relaxation effects.

3.
J Chem Phys ; 149(3): 034308, 2018 Jul 21.
Article in English | MEDLINE | ID: mdl-30037265

ABSTRACT

In this paper, we investigate HNCO by resonant and nonresonant Auger electron spectroscopy at the K-edges of carbon, nitrogen, and oxygen, employing soft X-ray synchrotron radiation. In comparison with the isosteric but linear CO2 molecule, spectra of the bent HNCO molecule are similar but more complex due to its reduced symmetry, wherein the degeneracy of the π-orbitals is lifted. Resonant Auger electron spectra are presented at different photon energies over the first core-excited 1s → 10a' resonance. All Auger electron spectra are assigned based on ab initio configuration interaction computations combined with the one-center approximation for Auger intensities and moment theory to consider vibrational motion. The calculated spectra were scaled by a newly introduced energy scaling factor, and generally, good agreement is found between experiment and theory for normal as well as resonant Auger electron spectra. A comparison of resonant Auger spectra with nonresonant Auger structures shows a slight broadening as well as a shift of the former spectra between -8 and -9 eV due to the spectating electron. Since HNCO is a small molecule and contains the four most abundant atoms of organic molecules, the reported Auger electron decay spectra will provide a benchmark for further theoretical approaches in the computation of core electron spectra.

4.
Anaesthesia ; 71(9): 1037-43, 2016 09.
Article in English | MEDLINE | ID: mdl-27523051

ABSTRACT

There are few data regarding postoperative hyperglycaemia in non-diabetic compared with diabetic patients following postoperative nausea and vomiting prophylaxis with dexamethasone. Eighty-five non-diabetic patients and patients with type-2 diabetes were randomly allocated to receive intravenous dexamethasone (8 mg) or ondansetron (4 mg). Blood glucose levels were measured at baseline and then 2, 4 and 24 h following induction of anaesthesia. In non-diabetic patients, the mean (SD) maximum blood glucose was higher in those who received dexamethasone compared with ondansetron (9.1 (2.2) mmol.l(-1) vs. 7.8 (1.4) mmol.l(-1) , p = 0.04). In diabetic patients, the mean (SD) maximum blood glucose was also higher in those who received dexamethasone compared with ondansetron (14.0 (2.5) mmol.l(-1) vs. 10.7 (2.4) mmol.l(-1) , p < 0.01). Multivariate analysis demonstrated that dexamethasone administration was a significant predictor of maximum postoperative blood glucose increase (p < 0.01) after adjusting for potential confounders. There was no interaction between baseline blood glucose level, or presence or absence of diabetes, and dexamethasone administration. We conclude that dexamethasone increases postoperative blood glucose levels in both non-diabetics and diabetics.


Subject(s)
Antiemetics/pharmacology , Blood Glucose/drug effects , Dexamethasone/pharmacology , Diabetes Mellitus, Type 2/blood , Postoperative Nausea and Vomiting/drug therapy , Adult , Dose-Response Relationship, Drug , Female , Humans , Male , Middle Aged , Ondansetron , Prospective Studies
5.
Nanotechnology ; 27(35): 355302, 2016 Sep 02.
Article in English | MEDLINE | ID: mdl-27454990

ABSTRACT

We employ Electron beam induced deposition (EBID) in combination with autocatalytic growth (AG) processes to fabricate magnetic nanostructures with controllable shapes and thicknesses. Following this route, different Fe deposits were prepared on silicon nitride membranes under ultra-high vacuum conditions and studied by scanning electron microscopy (SEM) and scanning transmission x-ray microspectroscopy (STXM). The originally deposited Fe nanostructures are composed of pure iron, especially when fabricated via autocatalytic growth processes. Quantitative near-edge x-ray absorption fine structure (NEXAFS) spectroscopy was employed to derive information on the thickness dependent composition. X-ray magnetic circular dichroism (XMCD) in STXM was used to derive the magnetic properties of the EBID prepared structures. STXM and XMCD analysis evinces the existence of a thin iron oxide layer at the deposit-vacuum interface, which is formed during exposure to ambient conditions. We were able to extract magnetic hysteresis loops for individual deposits from XMCD micrographs with varying external magnetic field. Within the investigated thickness range (2-16 nm), the magnetic coercivity, as evaluated from the width of the hysteresis loops, increases with deposit thickness and reaches a maximum value of ∼160 Oe at around 10 nm. In summary, we present a viable technique to fabricate ferromagnetic nanostructures in a controllable way and gain detailed insight into their chemical and magnetic properties.

6.
Nanoscale ; 6(7): 3695-703, 2014 Apr 07.
Article in English | MEDLINE | ID: mdl-24567084

ABSTRACT

This study addresses a combination of a well-developed and mild dispersion method and high-quality arc discharge single-walled carbon nanotubes (SWCNTs) as starting materials. Thus, we advance in fabrication of transparent, conducting films with extraordinary low material loss during SWCNT processing. The starting material was characterized by means of thermogravimetric analysis, high-resolution transmission electron microscopy and Raman spectroscopy. The quality of the starting material and produced dispersions was evaluated by ultraviolet and visible light absorption spectroscopy and Raman spectroscopy. A transparent conductive film was fabricated by drop-casting, whereas films were obtained with electrical to optical conductivity ratios (σDC/σOp) as high as 2.2, combined with a loss of nanotube material during processing well below 20 wt%. High pressure carbon monoxide conversion (HiPCO) SWCNTs, which are very well described in the literature, were used for comparison.

7.
J Chem Phys ; 140(2): 024503, 2014 Jan 14.
Article in English | MEDLINE | ID: mdl-24437892

ABSTRACT

Due to its importance for the function of organic optoelectronic devices, accurate simulations of the singlet exciton diffusion are crucial to predict the performance of new materials. We present a protocol which allows for the efficient directional analysis of exciton transport with high-level ab initio methods. It is based on an alternative to the frequently employed rate equation since the latter was found to be erroneous in some cases. The new approach can be used in combination with the master equation which is considerably faster than the corresponding Monte Carlo approach. The long-range character of the singlet exciton coupling is taken into account by an extrapolation scheme. The approach is applied to singlet exciton diffusion in those substances where these quantities are experimentally best established: naphthalene and anthracene. The high quality of the crystals, furthermore, diminish uncertainties arising from the geometrical structures used in the computations. For those systems, our new approach provides exciton diffusion lengths L for naphthalene and anthracene crystals which show an excellent agreement with their experimental counterparts. For anthracene, for example, the computed L value in a direction is computed to 58 nm while the experimental value is 60 ± 10 nm.

8.
Ann Oncol ; 24 Suppl 7: vii41-47, 2013 Oct.
Article in English | MEDLINE | ID: mdl-24001762

ABSTRACT

This article introduces palliative care to cancer patients in Middle Eastern countries. It considers the importance of the multidisciplinary team in providing an adequate service to the patient and his/her family. It provides views of professionals from the various countries with regard to the role of the nurse in such teams; whereby the three elements of palliative care nursing entail: 1. Working directly with patients and families; 2. Working with other health and social care professionals to network and co-ordinate services; and 3. working at an organizational level to plan, develop and manage service provision in local, regional and national settings. This article also details the challenges that nurses face in the Middle East and outlines the preferable ways to overcome such challenges. The latter include more focused educational activities at the undergraduate and graduate levels and continuous clinical training throughout their work as palliative care nurse specialists.


Subject(s)
Hospice and Palliative Care Nursing , Neoplasms/therapy , Patient Care Team , Aging , Humans , Middle East , Nurses , Palliative Care
9.
J Anim Sci ; 91(5): 2395-404, 2013 May.
Article in English | MEDLINE | ID: mdl-23463564

ABSTRACT

Antibiotics are used in ethanol production to control bacteria from competing with yeast for nutrients during starch fermentation. However, there is no published scientific information on whether antibiotic residues are present in distillers grains (DG), co-products from ethanol production, or whether they retain their biological activity. Therefore, the objectives of this study were to quantify concentrations of various antibiotic residues in DG and determine whether residues were biologically active. Twenty distillers wet grains and 20 distillers dried grains samples were collected quarterly from 9 states and 43 ethanol plants in the United States. Samples were analyzed for DM, CP, NDF, crude fat, S, P, and pH to describe the nutritional characteristics of the samples evaluated. Samples were also analyzed for the presence of erythromycin, penicillin G, tetracycline, tylosin, and virginiamycin M1, using liquid chromatography and mass spectrometry. Additionally, virginiamycin residues were determined, using a U.S. Food and Drug Administration-approved bioassay method. Samples were extracted and further analyzed for biological activity by exposing the sample extracts to 10(4) to 10(7) CFU/mL concentrations of sentinel bacterial strains Escherichia coli ATCC 8739 and Listeria monocytogenes ATCC 19115. Extracts that inhibited bacterial growth were considered to have biological activity. Physiochemical characteristics varied among samples but were consistent with previous findings. Thirteen percent of all samples contained low (≤1.12 mg/kg) antibiotic concentrations. Only 1 sample extract inhibited growth of Escherichia coli at 10(4) CFU/mL, but this sample contained no detectable concentrations of antibiotic residues. No extracts inhibited Listeria monocytogenes growth. These data indicate that the likelihood of detectable concentrations of antibiotic residues in DG is low; and if detected, they are found in very low concentrations. The inhibition in only 1 DG sample by sentinel bacteria suggests that antibiotic residues in DG were inactivated during the production process or are present in sublethal concentrations.


Subject(s)
Animal Feed/analysis , Anti-Bacterial Agents/pharmacology , Edible Grain/chemistry , Anti-Bacterial Agents/analysis , Chromatography, Liquid/veterinary , Colony Count, Microbial/veterinary , Escherichia coli/drug effects , Listeria monocytogenes/drug effects , Mass Spectrometry/veterinary , Minnesota
10.
J Chem Phys ; 137(4): 044310, 2012 Jul 28.
Article in English | MEDLINE | ID: mdl-22852621

ABSTRACT

High-resolution normal Auger-electron spectra of carbonyl sulfide subsequent to S 2p(-1) photoionization at photon energies of 200, 220, and 240 eV are reported along with corresponding photoelectron spectra. In addition, theoretical results are presented that take the core-hole orientation of the various spin-orbit-split and molecular-field-split S 2p(-1) states into account. Auger transitions to eight metastable dicationic final states are observed and assigned on the basis of the theoretical results. From Franck-Condon analysis, assuming Morse potentials along the normal coordinates for seven of the observed quasi-stable dicationic final states, information on the potential-energy surfaces is derived and compared with theoretical results from the literature.


Subject(s)
Quantum Theory , Sulfur Oxides/chemistry , Spectrum Analysis , Vibration
11.
J Appl Microbiol ; 113(5): 1076-86, 2012 Nov.
Article in English | MEDLINE | ID: mdl-22830299

ABSTRACT

AIMS: The objective of this study was to examine transcriptional changes in Escherichia coli when the bacterium was growing in the lettuce rhizoshpere. METHODS AND RESULTS: A combination of microarray analyses, colonization assays and confocal microscopy was used to gain a more complete understanding of bacterial genes involved in the colonization and growth of E. coli K12 in the lettuce root rhizosphere using a novel hydroponic assay system. After 3 days of interaction with lettuce roots, E. coli genes involved in protein synthesis, stress responses and attachment were up-regulated. Mutants in curli production (crl, csgA) and flagella synthesis (fliN) had a reduced capacity to attach to roots as determined by bacterial counts and by confocal laser scanning microscopy. CONCLUSIONS: This study indicates that E. coli K12 has the capability to colonize lettuce roots by using attachment genes and can readily adapt to the rhizosphere of lettuce plants. SIGNIFICANCE AND IMPACT OF THE STUDY: Results of this study show curli production and biofilm modulation genes are important for rhizosphere colonization and may provide useful targets to disrupt this process. Further studies using pathogenic strains will provide additional information about lettuce-E. coli interactions.


Subject(s)
Escherichia coli K12/genetics , Gene Expression Profiling , Lactuca/microbiology , Rhizosphere , Bacterial Adhesion , Escherichia coli K12/growth & development , Escherichia coli Proteins/genetics , Food Microbiology , Gene Expression Regulation, Bacterial , Gene Knockout Techniques , Genes, Bacterial , Hydroponics , Microscopy, Confocal , Oligonucleotide Array Sequence Analysis , Plant Roots/microbiology
12.
Nat Mater ; 11(6): 536-43, 2012 Apr 15.
Article in English | MEDLINE | ID: mdl-22504534

ABSTRACT

Molecular orientation critically influences the mechanical, chemical, optical and electronic properties of organic materials. So far, molecular-scale ordering in soft matter could be characterized with X-ray or electron microscopy techniques only if the sample exhibited sufficient crystallinity. Here, we show that the resonant scattering of polarized soft X-rays (P-SoXS) by molecular orbitals is not limited by crystallinity and that it can be used to probe molecular orientation down to size scales of 10 nm. We first apply the technique on highly crystalline small-molecule thin films and subsequently use its high sensitivity to probe the impact of liquid-crystalline ordering on charge mobility in polymeric transistors. P-SoXS also reveals scattering anisotropy in amorphous domains of all-polymer organic solar cells where interfacial interactions pattern orientational alignment in the matrix phase, which probably plays an important role in the photophysics. The energy and q-dependence of the scattering anisotropy allows the identification of the composition and the degree of orientational order in the domains.

13.
Br J Pharmacol ; 165(5): 1467-75, 2012 Mar.
Article in English | MEDLINE | ID: mdl-21838752

ABSTRACT

BACKGROUND AND PURPOSE: TASK1 (K(2P)3.1) two-pore-domain K(+) channels contribute substantially to the resting membrane potential in human pulmonary artery smooth muscle cells (hPASMC), modulating vascular tone and diameter. The endothelin-1 (ET-1) pathway mediates vasoconstriction and is an established target of pulmonary arterial hypertension (PAH) therapy. ET-1-mediated inhibition of TASK1 currents in hPASMC is implicated in the pathophysiology of PAH. This study was designed to elucidate molecular mechanisms underlying inhibition of TASK1 channels by ET-1. EXPERIMENTAL APPROACH: Two-electrode voltage clamp and whole-cell patch clamp electrophysiology was used to record TASK1 currents from hPASMC and Xenopus oocytes. KEY RESULTS: ET-1 inhibited TASK1-mediated I(KN) currents in hPASMC, an effect attenuated by Rho kinase inhibition with Y-27632. In Xenopus oocytes, TASK1 current reduction by ET-1 was mediated by endothelin receptors ET(A) (IC(50) = 0.08 nM) and ET(B) (IC(50) = 0.23 nM) via Rho kinase signalling. TASK1 channels contain two putative Rho kinase phosphorylation sites, Ser(336) and Ser(393) . Mutation of Ser(393) rendered TASK1 channels insensitive to ET(A) - or ET(B)-mediated current inhibition. In contrast, removal of Ser(336) selectively attenuated ET(A) -dependent TASK1 regulation without affecting the ET(B) pathway. CONCLUSIONS AND IMPLICATIONS: ET-1 regulated vascular TASK1 currents through ET(A) and ET(B) receptors mediated by downstream activation of Rho kinase and direct channel phosphorylation. The Rho kinase pathway in PASMC may provide a more specific therapeutic target in pulmonary arterial hypertension treatment.


Subject(s)
Endothelin-1/metabolism , Nerve Tissue Proteins/antagonists & inhibitors , Potassium Channels, Tandem Pore Domain/antagonists & inhibitors , rho-Associated Kinases/metabolism , Animals , Cells, Cultured , Female , GTP Phosphohydrolases/metabolism , Humans , Hypertension, Pulmonary/genetics , Hypertension, Pulmonary/metabolism , Hypertension, Pulmonary/physiopathology , Membrane Potentials/genetics , Membrane Potentials/physiology , Muscle, Smooth, Vascular/metabolism , Muscle, Smooth, Vascular/physiology , Mutation , Myocytes, Smooth Muscle/metabolism , Myocytes, Smooth Muscle/physiology , Nerve Tissue Proteins/genetics , Nerve Tissue Proteins/metabolism , Phosphorylation , Potassium Channels, Tandem Pore Domain/genetics , Potassium Channels, Tandem Pore Domain/metabolism , Pulmonary Artery/metabolism , Pulmonary Artery/physiology , Receptor, Endothelin A/metabolism , Receptor, Endothelin B/metabolism , Signal Transduction , Vasoconstriction/genetics , Vasoconstriction/physiology , Xenopus laevis , rho-Associated Kinases/antagonists & inhibitors
14.
Phys Chem Chem Phys ; 13(41): 18428-35, 2011 Nov 07.
Article in English | MEDLINE | ID: mdl-21922100

ABSTRACT

Multi-electron coincidence measurements on photoionisation of H(2)S have been carried out at photon energies from 40 to 250 eV. They quantify molecular field effects on the Auger process in detail and are in good agreement with the existing theory. Spectra of core-valence double ionisation of H(2)S are presented and partially analysed. Auger decays from the core-valence states produce triply charged product spectra with unexplained and surprising intensity distributions. Triple ionisation by the double Auger process from 2p hole states shows little effect of the molecular field splitting, but includes a substantial contribution from cascade processes, some involving dissociation in intermediate states. The onset of triple ionisation at the molecular geometry is determined as 61 ± 0.5 eV.

15.
Br J Anaesth ; 106(6): 851-5, 2011 Jun.
Article in English | MEDLINE | ID: mdl-21531744

ABSTRACT

Remifentanil undergoes extensive placental transfer and has been used to provide fetal immobilization and anaesthesia for in utero fetal endoscopic interventions. We report three cases of the ex utero intrapartum treatment performed under neuraxial anaesthesia where the maternal administration of remifentanil was used to provide fetal immobilization and analgesia. Fetal pathology included goiter and arthrogryposis, with one case requiring a tracheostomy. The longest time on placental circulation was 21 min. No clinically significant maternal sedation or respiratory depression was observed. In all cases, remifentanil provided adequate fetal immobilization and obviated the need to administer other analgesics or neuromuscular blocking agents. Remifentanil is a useful adjunct for ex utero fetal procedures.


Subject(s)
Anesthesia, Obstetrical/methods , Fetal Movement/drug effects , Fetal Therapies/methods , Hypnotics and Sedatives/pharmacology , Piperidines/pharmacology , Adult , Anesthesia, Epidural , Anesthesia, Spinal , Arthrogryposis/surgery , Female , Fetal Diseases/surgery , Goiter/surgery , Humans , Immobilization/methods , Peripartum Period , Pregnancy , Remifentanil , Young Adult
16.
Phys Chem Chem Phys ; 13(20): 9839-48, 2011 May 28.
Article in English | MEDLINE | ID: mdl-21503316

ABSTRACT

The molecular interaction of dihydroxo[5,10,15,20-tetrakis(4-tert-butyl-phenyl)porphyrinato]-tin(IV) (SnTTBPP(OH)(2)), the structural order and growth of ultrathin films on Ag(100) have been studied by means of low-energy electron diffraction (LEED) and synchrotron based photoelectron spectroscopy, i.e., X-ray photoemission (XPS) and near-edge X-ray absorption fine structure (NEXAFS/XANES) spectroscopy. For the first time, monolayer adsorption of a metalloporphyrin with octahedral coordination of the metal center by two additional axial hydroxo ligands is investigated in a multi-technique study. The delicate balance of molecule-substrate interactions and intermolecular interactions leads to the formation of a densely-packed organic monolayer which is commensurate with the Ag(100) substrate. From NEXAFS linear dichroism an almost coplanar orientation of the porphyrin system is derived. XPS and NEXAFS clearly indicate that the axial hydroxo ligands are cleaved in monolayer films, i.e., upon adsorption to the Ag substrate. With increasing film thickness orientational order gets lost and leads to polycrystalline growth for thicker films as confirmed by scanning X-ray transmission microscopy (STXM).


Subject(s)
Organotin Compounds/chemistry , Silver/chemistry , Tin/chemistry , Adsorption , Metalloporphyrins/chemistry , Organotin Compounds/chemical synthesis , X-Ray Absorption Spectroscopy
17.
Ultramicroscopy ; 110(11): 1358-61, 2010 Oct.
Article in English | MEDLINE | ID: mdl-20692099

ABSTRACT

The lateral resolution of a surface sensitive low-energy electron microscope (LEEM) has been improved below 4 nm for the first time. This breakthrough has only been possible by simultaneously correcting the unavoidable spherical and chromatic aberrations of the lens system. We present an experimental criterion to quantify the aberration correction and to optimize the electron optical system. The obtained lateral resolution of 2.6 nm in LEEM enables the first surface sensitive, electron microscopic observation of the herringbone reconstruction on the Au(111) surface.

18.
Science ; 329(5989): 303-5, 2010 Jul 16.
Article in English | MEDLINE | ID: mdl-20647462

ABSTRACT

Inverse melting or disordering, in which the disordered phase forms upon cooling, is known for a few cases in bulk systems under high pressure. We show that inverse disordering also occurs in two dimensions: For a monolayer of 1,4,5,8-naphthalene-tetracarboxylic dianhydride on Ag(111), a completely reversible order-disorder transition appears upon cooling. The transition is driven by strongly anisotropic interactions within the layer versus with the metal substrate. Spectroscopic data reveal changes in the electronic structure of the system corresponding to a strengthening of the interface bonding at low temperatures. We demonstrate that the delicate, temperature-dependent balance between the vertical and lateral forces is the key to understanding this unconventional phase transition.

19.
Rev Sci Instrum ; 81(3): 033704, 2010 Mar.
Article in English | MEDLINE | ID: mdl-20370182

ABSTRACT

The successful integration of electron detection into an existing scanning transmission x-ray microspectroscope (STXM) at the Swiss Light Source is demonstrated. In conventional x-ray detection using a photomultiplier, STXM offers mainly bulk sensitivity combined with high lateral resolution. However, by implementation of a channeltron electron multiplier, the surface sensitivity can be established by the detection of secondary electrons emitted from the sample upon resonant excitation. We describe the experimental setup and discuss several relevant aspects, in particular the schemes to correct for self-absorption in the specimen due to back illumination in case of thicker films.

20.
Biophys J ; 98(4): 606-16, 2010 Feb 17.
Article in English | MEDLINE | ID: mdl-20159157

ABSTRACT

Progressive force loss in Duchenne muscular dystrophy is characterized by degeneration/regeneration cycles and fibrosis. Disease progression may involve structural remodeling of muscle tissue. An effect on molecular motorprotein function may also be possible. We used second harmonic generation imaging to reveal vastly altered subcellular sarcomere microarchitecture in intact single dystrophic mdx muscle cells (approximately 1 year old). Myofibril tilting, twisting, and local axis deviations explain at least up to 20% of force drop during unsynchronized contractile activation as judged from cosine angle sums of myofibril orientations within mdx fibers. In contrast, in vitro motility assays showed unaltered sliding velocities of single mdx fiber myosin extracts. Closer quantification of the microarchitecture revealed that dystrophic fibers had significantly more Y-shaped sarcomere irregularities ("verniers") than wild-type fibers (approximately 130/1000 microm(3) vs. approximately 36/1000 microm(3)). In transgenic mini-dystrophin-expressing fibers, ultrastructure was restored (approximately 38/1000 microm(3) counts). We suggest that in aged dystrophic toe muscle, progressive force loss is reflected by a vastly deranged micromorphology that prevents a coordinated and aligned contraction. Second harmonic generation imaging may soon be available in routine clinical diagnostics, and in this work we provide valuable imaging tools to track and quantify ultrastructural worsening in Duchenne muscular dystrophy, and to judge the beneficial effects of possible drug or gene therapies.


Subject(s)
Molecular Motor Proteins/chemistry , Molecular Motor Proteins/metabolism , Muscle, Skeletal/metabolism , Muscular Dystrophies/metabolism , Aging , Algorithms , Animals , Biomechanical Phenomena , Dystrophin/metabolism , Gene Expression Regulation , Humans , Imaging, Three-Dimensional , Mice , Mice, Inbred mdx , Mice, Transgenic , Microscopy , Molecular Imaging , Movement , Muscle Fibers, Skeletal/metabolism , Muscle, Skeletal/pathology , Muscle, Skeletal/physiology , Muscular Dystrophies/physiopathology , Sarcomeres/metabolism
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