JoVE   
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Biology

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Neuroscience

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Immunology and Infection

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Clinical and Translational Medicine

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Bioengineering

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Applied Physics

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Chemistry

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Behavior

  
You do not have subscription access to articles in this section. Learn more about access.

  JoVE Environment

|   

JoVE Science Education

General Laboratory Techniques

You do not have subscription access to videos in this collection. Learn more about access.

Basic Methods in Cellular and Molecular Biology

You do not have subscription access to videos in this collection. Learn more about access.

Model Organisms I

You do not have subscription access to videos in this collection. Learn more about access.

Model Organisms II

You do not have subscription access to videos in this collection. Learn more about access.

Essentials of
Neuroscience

You do not have subscription access to videos in this collection. Learn more about access.

Essentials of Developmental Biology

You have subscription access to videos in this collection through your user account.

In JoVE (1)

Other Publications (200)

Articles by Adrien E. Schramm in JoVE

 JoVE Neuroscience

Targeted Labeling of Neurons in a Specific Functional Micro-domain of the Neocortex by Combining Intrinsic Signal and Two-photon Imaging

1Department of Neuroscience, Medical University of South Carolina


JoVE 50025

A method is described for labeling neurons with fluorescent dyes in predetermined functional micro-domains of the neocortex. First, intrinsic signal optical imaging is used to obtain a functional map. Then two-photon microscopy is used to label and image neurons within a micro-domain of the map.

Other articles by Adrien E. Schramm on PubMed

Development of Inhibitors in Haemophilia. Ongoing Epidemiological Study

A number of 513 consecutive patients (494-haemophilia A and 19-haemophilia B) from eight haemophilia treatment centers have been investigated with Bethesda assay for the presence of factor VIII or IX inhibitors. The overall prevalence of inhibitors was 15.20%, 18.48% in severe, 5.60% in moderate and 12.24% in mild forms. The prevalence was higher than reported in most of the western countries. The age at start of substitution (p = 0.9775), the frequent switching of factor concentrates (p = 0.8931) were not relevant factors for the development of inhibitors. It is worth to be mentioned the unexpectedly occurrence of inhibitors in prior inhibitor negative (6/72) patients (during surgical interventions) probably due to their previous scarce substitution, occurrence which seems not being connected with the continuous infusion modality of factor VIII administration (p = 0.8341). In controversial situations, in the field of low titer (≤ 1 BU/ml) inhibitors for a reliable interpretation of the results the performance of recovery index and half-life time assessment of FVIII/IX was undertaken.

Haemophilia Patients with HIV- and HCV-infection. Long Term Survival

At present (2010), we investigate in a long term study on psychosocial aspects of haemophilia at the Munich Haemophilia Centre factors which could be held responsible for the large number of still living HIV+ patients while many patients died from AIDS or from diseases caused by HCV during the 1990ies. PATIENTS, METHODS: We retrospectively compare medical and psychosocial data of 15 HIV+ long term survivors (L-S), 15 HIV+ later on deceased patients and 15 HIV-, all of them deriving from the original investigation group from 1985 including 52 patients (30 HIV+, 22 HIV-). We prove the validity of our psychosocial factors by the SF 36 which currently serves as gold standard. The actual psychosocial factors and medical parameters of the 15 L-S are checked against the data of the 15 HIV- patients. Results, conclusion: In 1985 already, we retrospectively found significantly higher values regarding psychosocial factors within the group of the L-S as opposed to the later on deceased ones. Highly significant are the facts that more than 90 % of HIV+ L-S have had a good relationship to their fathers and are still employed today in contrast to the deceased HIV+ patients. At present, the differences regarding psychosocial factors between HIV+ L-S and HIV-negative patients are vanishing.

Mass-dependent Bond Vibrational Dynamics Influence Catalysis by HIV-1 Protease

Protein motions that occur on the microsecond to millisecond time scale have been linked to enzymatic rates observed for catalytic turnovers, but not to transition-state barrier crossing. It has been hypothesized that enzyme motions on the femtosecond time scale of bond vibrations play a role in transition state formation. Here, we perturb femtosecond motion by substituting all nonexchangeable carbon, nitrogen, and hydrogen atoms with (13)C, (15)N, and (2)H and observe the catalytic effects in HIV-1 protease. According to the Born-Oppenheimer approximation, isotopic substitution alters vibrational frequency with unchanged electrostatic properties. With the use of a fluorescent peptide to report on multiple steps in the reaction, we observe significantly reduced rates in the heavy enzyme relative to the light enzyme. A possible interpretation of our results is that there exists a dynamic link between mass-dependent bond vibrations of the enzyme and events in the reaction coordinate.

Femtosecond Dynamics Coupled to Chemical Barrier Crossing in a Born-Oppenheimer Enzyme

Contributions of fast (femtosecond) dynamic motion to barrier crossing at enzyme catalytic sites is in dispute. Human purine nucleoside phosphorylase (PNP) forms a ribocation-like transition state in the phosphorolysis of purine nucleosides and fast protein motions have been proposed to participate in barrier crossing. In the present study, (13)C-, (15)N-, (2)H-labeled human PNP (heavy PNP) was expressed, purified to homogeneity, and shown to exhibit a 9.9% increase in molecular mass relative to its unlabeled counterpart (light PNP). Kinetic isotope effects and steady-state kinetic parameters were indistinguishable for both enzymes, indicating that transition-state structure, equilibrium binding steps, and the rate of product release were not affected by increased protein mass. Single-turnover rate constants were slowed for heavy PNP, demonstrating reduced probability of chemical barrier crossing from enzyme-bound substrates to enzyme-bound products. In a second, independent method to probe barrier crossing, heavy PNP exhibited decreased forward commitment factors, also revealing mass-dependent decreased probability for barrier crossing. Increased atomic mass in human PNP alters bond vibrational modes on the femtosecond time scale and reduces on-enzyme chemical barrier crossing. This study demonstrates coupling of enzymatic bond vibrations on the femtosecond time scale to barrier crossing.

Whipple Limbic Encephalitis

To describe a relapse of Whipple disease revealed by isolated limbic encephalitis with no other signs of systemic involvement.

Clinical and Radiographic Evaluation of Early Loaded Narrow Diameter Implants - 1-year Follow-up

OBJECTIVE: To evaluate the clinical performance of Astra Tech OsseoSpeed™ TX 3.0S implants using one-stage surgical procedure and early loading in the anterior region. MATERIAL AND METHODS: This is a prospective, single arm, multi-centre study. Patients missing teeth at positions 12, 22 and 32-42 were eligible to enter the study. The implants (OsseoSpeed™ TX 3.0S) used in the study were of 3 mm diameter and of different lengths. One-stage surgery was performed, and healing abutments were used during the 6-10 weeks healing period. Clinical and radiographic examinations were assessed at implant installation, loading and at the 6- and 12-month follow-up visits. RESULTS: Ninety-seven implants were placed in 69 patients at six different study centres in Denmark, Germany, Italy, Spain, Sweden and the United Kingdom. The survival rate was 95.9%. No implants have been lost after loading (100% survival rate after loading). Mean marginal bone loss 1 year after installation was 0.065 mm (SD = 1.018). The frequency of bone loss ≥1 mm was 6.6% and 51.3% of the implants demonstrated no bone loss or even bone gain from the surgical visit to the first year follow-up visit. Mean probing pocket depth and gingival zenith score were stable from crown placement to the 6- and 1-year follow-up visits. CONCLUSION: Treatment with OsseoSpeed™ TX 3.0S implants is a safe and predictable option in the anterior region where physical space is limited. Minimal marginal bone loss was observed during the first year follow-up.

Detection of Neuroblastoma Cells During Clinical Follow Up: Advanced Flow Cytometry and Rt-PCR for Tyrosine Hydroxylase Using Both Conventional and Real-time PCR

Real-time reverse-transcriptase PCR (RT-qPCR) or conventional RT-PCR (RT-cPCR) detection of tyrosine hydroxylase (TH) is increasingly used to detect neuroblastoma (NB) cells in clinical samples. However, TH expression in normal tissues can limit its usefulness and make additional diagnostic strategies necessary.

Plasmodium Falciparum Parasites Are Killed by a Transition State Analogue of Purine Nucleoside Phosphorylase in a Primate Animal Model

Plasmodium falciparum causes most of the one million annual deaths from malaria. Drug resistance is widespread and novel agents against new targets are needed to support combination-therapy approaches promoted by the World Health Organization. Plasmodium species are purine auxotrophs. Blocking purine nucleoside phosphorylase (PNP) kills cultured parasites by purine starvation. DADMe-Immucillin-G (BCX4945) is a transition state analogue of human and Plasmodium PNPs, binding with picomolar affinity. Here, we test BCX4945 in Aotus primates, an animal model for Plasmodium falciparum infections. Oral administration of BCX4945 for seven days results in parasite clearance and recrudescence in otherwise lethal infections of P. falciparum in Aotus monkeys. The molecular action of BCX4945 is demonstrated in crystal structures of human and P. falciparum PNPs. Metabolite analysis demonstrates that PNP blockade inhibits purine salvage and polyamine synthesis in the parasites. The efficacy, oral availability, chemical stability, unique mechanism of action and low toxicity of BCX4945 demonstrate potential for combination therapies with this novel antimalarial agent.

Electron Temperature Scaling in Laser Interaction with Solids

A precise knowledge of the temperature and number of hot electrons generated in the interaction of short-pulse high-intensity lasers with solids is crucial for harnessing the energy of a laser pulse in applications such as laser-driven ion acceleration or fast ignition. Nevertheless, present scaling laws tend to overestimate the hot electron temperature when compared to experiment and simulations. We present a novel approach that is based on a weighted average of the kinetic energy of an ensemble of electrons. We find that the scaling of electron energy with laser intensity can be derived from a general Lorentz invariant electron distribution ansatz that does not rely on a specific model of energy absorption. The scaling derived is in perfect agreement with simulation results and clearly follows the trend seen in recent experiments, especially at high laser intensities where other scalings fail to describe the simulations accurately.

Dragon PolyA Spotter: Predictor of Poly(A) Motifs Within Human Genomic DNA Sequences

MOTIVATION: Recognition of poly(A) signals in mRNA is relatively straightforward due to the presence of easily recognizable polyadenylic acid tail. However, the task of identifying poly(A) motifs in the primary genomic DNA sequence that correspond to poly(A) signals in mRNA is a far more challenging problem. Recognition of poly(A) signals is important for better gene annotation and understanding of the gene regulation mechanisms. In this work, we present one such poly(A) motif prediction method based on properties of human genomic DNA sequence surrounding a poly(A) motif. These properties include thermodynamic, physico-chemical and statistical characteristics. For predictions, we developed Artificial Neural Network and Random Forest models. These models are trained to recognize 12 most common poly(A) motifs in human DNA. Our predictors are available as a free web-based tool accessible at http://cbrc.kaust.edu.sa/dps. Compared with other reported predictors, our models achieve higher sensitivity and specificity and furthermore provide a consistent level of accuracy for 12 poly(A) motif variants. CONTACT: vladimir.bajic@kaust.edu.sa SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Osmotic Repulsion Force Due to Adsorbed Surfactants

When considering interaction forces in surfactant-stabilized colloidal dispersions a factor that has been rarely discussed is the possible effect of osmotic force due to overlapping adsorbed surfactant monolayers. In the present work, the osmotic repulsion force is built-in on the basis of DLVO mechanics and based on Fischer's consideration of the analogous situation for adsorbed polymer layers on solid surfaces [E.W. Fischer, Kolloid Zeitschrift 160 (1958) 120-141] and on Langmuir's earlier concept of osmotic pressure excess due to overlapping adsorption layers [I. Langmuir, J. Chem. Phys. 6 (1938) 873-896]. The advanced method for calculation of the net repulsion force in overlapping surfactant monolayers is developed and applied to real adsorbed surfactant systems. We show that the value of disjoining pressure can reach values as high as 8 MPa for the condition of fully overlapping surfactant adsorption layers, based on the calculation of the first virial term of the general expression for osmotic pressure. Thus, we have shown that osmotic forces can be substantial at distances of close interfacial approach, and that they can easily be of the same or greater order of magnitude than the forces that have been more conventionally considered.

Seasonal Variability in Cadmium, Lead, Copper, Zinc and Iron Concentrations in the Three Major Fish Species, Oreochromis Niloticus, Lates Niloticus and Rastrineobola Argentea in Winam Gulf, Lake Victoria: Impact of Wash-off into the Lake

Trace metals Cadmium (Cd), Lead (Pb), Copper (Cu), Zinc (Zn) and Iron (Fe) were analyzed in edible portions of three main finfish species namely Lates niloticus, Oreochromis niloticus and Rastrineobola argentea sampled from various beaches of Winam Gulf, Lake Victoria, Kenya, in order to determine any seasonal and site variations and the results showed significantly (p < 0.05) higher mean concentrations of Cd, Cu, Zn and Fe during the wet season compared to the dry season for all the three species indicating the impact of wash-off into the lake during the rainy periods. The overall mean concentrations of the heavy metals (in μg/g dry weight) in all combined samples ranged from 0.17-0.40 (Cd), 0.47-2.53 (Pb), 2.13-8.74 (Cu), 28.9-409.3 (Zn) and 31.4-208.1 (Fe), respectively. It was found that consumption of Rastrineobola argentea can be a significant source of heavy metals especially Zn, to humans, compared with Lates niloticus and Oreochromis niloticus, if only the muscle parts of the latter two are consumed.

Clinical Features Cannot Distinguish Allergic from Non-allergic Asthma in Children

Environmental allergens are a major trigger of asthma, but not all asthmatics are allergic. This study was designed to review clinical characteristics in children with allergic and non-allergic asthma, based on responsiveness to allergy skin tests, in order to identify a combination of features that could distinguish allergic from non-allergic asthma in children.

Laboratory Studies on Formation and Minimisation of Polychlorinated Dibenzodioxins and -furans (PCDD/F) in Secondary Aluminium Process

The objectives of this work were to study the formation mechanisms of polychlorinated dibenzo-p-dioxins/polychlorinated dibenzofurans (PCDD/F) in thermal aluminium recycling processes by use of laboratory experiments. The pattern of isomers of PCDD/F indicates that de novo synthesis is important in aluminium smeltery. The mechanisms of PCDD/F formation in aluminium smelting are similar to that of various incineration processes of waste material. The results of bioanalysis (EROD-test) confirms the existence of de novo synthesis of PCDD/F, but points out to the existence to some additional, toxic compounds of unknown structure. To reduce the amount of PCDD/F the input of carbon at the metal should be reduced; in addition the metal smeltery plants should be cleaned from fly ash particles. It is suggested to use good primary methods in the technical plants like constant feeding of the metal into the oven will minimise PCDD/F concentration. The biological EROD-bioassay is a good tool to estimate PCDD/F-TEQ values also for this technical process simulated in the laboratory.

Human and Environmental Biomonitoring of Polychlorinated Biphenyls and Hexachlorobenzene in Saxony, Germany Based on the German Environmental Specimen Bank

The objective of the present study was to investigate the principle relationships between concentrations in human and environmental matrices of polychlorinated biphenyls (PCBs) and hexachlorobenzene (HCB) in short distance comparable areas within Saxony, Germany by employing the data of the German Environmental Specimen Banking (ESB). Examples supporting this idea were presented by selecting data on blood plasma collected from students in University of Halle and pine shoots, egg matter of city pigeons, earthworm, and roe deer liver. Similar pattern for PCB 138 and PCB 180 was found for the human plasma and pine shoots samples during investigated years and the human data followed the corresponding environmental levels with some delay of approximately two years. However, PCB 153 that was the prevailing congener did not manifest this relationship. In addition, the correlation of the ratios of concentrations (human/environmental concentration) to some physicochemical constants such as molecular weight (MW), octanol-water partition coefficient (logK(ow)), Henry's law constant (K(H)), and sorption partition coefficient (logK(oc)) of HCB, PCB 138, PCB 153, and PCB 180 were studied. The resulted negative slopes with all constants in case of blood plasma/city pigeons egg matter pairs suggested that the accumulation of lipophilic compounds is more pronounced in pigeon eggs than in human blood.

Investigating the Biocompatibility of Two New Heavy Intraocular Dyes for Vitreoretinal Surgery with an Isolated Perfused Vertebrate Retina Organ Culture Model and a Retinal Ganglion Cell Line

During vitreoretinal surgery, vital dyes are used to visualize anatomical structures. Substances with a density higher than water are added to facilitate sedimentation and staining. BBG with 4% PEG (ILM Blue) and BBG with TB and 4% PEG (MBB Dual) are two new dyes. This study evaluates biocompatibility of the new dye solutions, using cell cultures and electrophysiological evaluation.

Aberrations of the Cathode Objective Lens Up to Fifth Order

In this paper we discuss a topic that was close to Prof. Gertrude Rempfer s interests for many years. On this occasion of her 100th birthday, we remember and honor Gertrude for her many outstanding contributions, and for the inspiring example that she set. We derive theoretical expressions for the aberration coefficients of the uniform electrostatic field up to 5th order and compare these with raytracing calculations for the cathode lens used in Low Energy Electron Microscopy and Photo Electron Emission Microscopy experiments. These higher order aberration coefficients are of interest for aberration corrected experiments in which chromatic (C(c)) and spherical (C₃) aberrations of the microscope are set to zero. The theoretical predictions are in good agreement with the results of raytracing. Calculations of image resolution using the Contrast Transfer Function method show that sub-nanometer resolution is achievable in an aberration corrected LEEM system.

Metabolism of Glucosinolate-derived Isothiocyanates to Glutathione Conjugates in Generalist Lepidopteran Herbivores

The defensive properties of the glucosinolate-myrosinase system in plants of the order Brassicales have been attributed to the formation of toxic isothiocyanates generated upon tissue damage. Lepidopteran herbivores specialised on brassicaceous plants have been shown to possess biochemical mechanisms preventing the formation of isothiocyanates. Yet, no such mechanisms are known for generalist lepidopterans which also occasionally but successfully feed on plants of the Brassicales. After feeding on Arabidopsis thaliana plants, faeces of Spodoptera littoralis larvae contained glutathione conjugate derivatives (cysteinylglycine- and cysteinyl-isothiocyanate-conjugates) of the plant's major glucosinolate hydrolysis product, 4-methylsulfinylbutyl isothiocyanate. When caterpillars fed on leaves of A. thaliana containing [¹⁴C]₄-methylsulfinylbutyl glucosinolate, more than half of the ingested radioactivity was excreted as the unmetabolised corresponding isothiocyanate, and only 11% as glutathione conjugate derivatives. However, these conjugates were demonstrated to be the major metabolites of isothiocyanates in S. littoralis, and their abundance was shown to correlate with the amount of isothiocyanates ingested. Analysis of larval faeces from several species of generalist lepidopterans (Spodoptera exigua, S. littoralis, Mamestra brassicae, Trichoplusia ni and Helicoverpa armigera) fed on different Brassicaceae revealed that glutathione conjugates arise from a variety of aliphatic and aromatic isothiocyanates derived from dietary glucosinolates.

Studies on Differential Nuclear Translocation Mechanism and Assembly of the Three Subunits of the Arabidopsis Thaliana Transcription Factor NF-Y

The eukaryotic transcription factor NF-Y consists of three subunits (A, B, and C), which are encoded in Arabidopsis thaliana in multigene families consisting of 10, 13, and 13 genes, respectively. In principle, all potential combinations of the subunits are possible for the assembly of the heterotrimeric complex. We aimed at assessing the probability of each subunit to participate in the assembly of NF-Y. The evaluation of physical interactions among all members of the NF-Y subunit families indicate a strong requirement for NF-YB/NF-YC heterodimerization before the entire complex can be accomplished. By means of a modified yeast two-hybrid system assembly of all three subunits to a heterotrimeric complex was demonstrated. Using GFP fusion constructs, NF-YA and NF-YC localization in the nucleus was demonstrated, while NF-YB is solely imported into the nucleus as a NF-YC-associated heterodimer NF-YC. This piggyback transport of the two Arabidopsis subunits differs from the import of the NF-Y heterotrimer of heterotrophic organisms. Based on a peptide structure model of the histone-fold-motifs, disulfide bonding among intramolecular conserved cysteine residues of NF-YB, which is responsible for the redox-regulated assembly of NF-YB and NF-YC in human and Aspergillus nidulans, can be excluded for Arabidopsis NF-YB.

The Cross-linked Biopolymer Hyaluronic Acid As an Artificial Vitreous Substitute

Biopolymers are promising substances in the development of a new vitreous substitute to overcome the drawbacks associated with current hydrophobic tamponade materials.

A Contrast Transfer Function Approach for Image Calculations in Standard and Aberration-corrected LEEM and PEEM

We introduce an extended Contrast Transfer Function (CTF) approach for the calculation of image formation in low energy electron microscopy (LEEM) and photo electron emission microscopy (PEEM). This approach considers aberrations up to fifth order, appropriate for image formation in state-of-the-art aberration-corrected LEEM and PEEM. We derive Scherzer defocus values for both weak and strong phase objects, as well as for pure amplitude objects, in non-aberration-corrected and aberration-corrected LEEM. Using the extended CTF formalism, we calculate contrast and resolution of one-dimensional and two-dimensional pure phase, pure amplitude, and mixed phase and amplitude objects. PEEM imaging is treated by adapting this approach to the case of incoherent imaging. Based on these calculations, we show that the ultimate resolution in aberration-corrected LEEM is about 0.5 nm, and in aberration-corrected PEEM about 3.5 nm. The aperture sizes required to achieve these ultimate resolutions are precisely determined with the CTF method. The formalism discussed here is also relevant to imaging with high resolution transmission electron microscopy.

Transcatheter Aortic Valve Implantation in Aortic Coarctation

A 77-year-old male patient was scheduled for transcatheter aortic valve implantation for symptomatic and severe aortic valve stenosis. Severe multidirectional kinking of the aorta based on aortic coarctation did not allow for the transfemoral, but only for the transapical approach. The procedure was complicated because of the technically challenging retrograde passage of the transfemorally inserted pig-tail catheter required for intraoperative angiography of the aortic root. Correct positioning of the pig-tail catheter into the ascending aorta was accomplished by use of a loop snare, which was advanced into the descending aorta via the antegrade route, passing the cardiac apex, the stenotic aortic valve, and the coarctation-associated kinking. The pig-tail catheter tip was manipulated into the loop snare, pulled traverse the coarctation, and released within the proximal ascending aorta. Subsequent procedures were uneventful and followed the standardized protocol. A 29 mm Edwards Lifescience transcatheter Sapien bioprosthesis was successfully implanted.

A Complex of Methylthioadenosine/S-adenosylhomocysteine Nucleosidase, Transition State Analogue, and Nucleophilic Water Identified by Mass Spectrometry

An enzyme-stabilized nucleophilic water molecule has been implicated at the transition state of Escherichia coli methylthioadenosine nucleosidase (EcMTAN) by transition state analysis and crystallography. We analyzed the EcMTAN mass in complex with a femtomolar transition state analogue to determine whether the inhibitor and nucleophilic water could be detected in the gas phase. EcMTAN-inhibitor and EcMTAN-inhibitor-nucleophilic water complexes were identified by high-resolution mass spectrometry under nondenaturing conditions. The enzyme-inhibitor-water complex is sufficiently stable to exist in the gas phase.

Expression Profiles of Human Interferon-alpha and Interferon-lambda Subtypes Are Ligand- and Cell-dependent

Recent genome-wide association studies suggest distinct roles for 12 human interferon-alpha (IFN-α) and 3 IFN-λ subtypes that may be elucidated by defining the expression patterns of these sets of genes. To overcome the impediment of high homology among each of the sets, we designed a quantitative real-time PCR assay that incorporates the use of molecular beacon and locked nucleic acid (LNA) probes, and in some instances, LNA oligonucleotide inhibitors. We then measured IFN subtype expression by human peripheral blood mononuclear cells and by purified monocytes, myeloid dendritic cells (mDC), plasmacytoid dendritic cells (pDC), and monocyte-derived macrophages (MDM), and -dendritic cells (MDDC) in response to poly I:C, lipopolysaccharide (LPS), imiquimod and CpG oligonucleotides. We found that in response to poly I:C and LPS, monocytes, MDM and MDDC express a subtype pattern restricted primarily to IFN-β and IFN-λ1. In addition, while CpG elicited expression of all type I IFN subtypes by pDC, imiquimod did not. Furthermore, MDM and mDC highly express IFN-λ, and the subtypes of IFN-λ are expressed hierarchically in the order IFN-λ1 followed by IFN-λ2, and then IFN-λ3. These data support a model of coordinated cell- and ligand-specific expression of types I and III IFN. Defining IFN subtype expression profiles in a variety of contexts may elucidate specific roles for IFN subtypes as protective, therapeutic or pathogenic mediators.

The Metallophosphodiesterase Mpped2 Impairs Tumorigenesis in Neuroblastoma

Through microarray analyses, we identified the Mpped2 gene as differentially expressed in two neuroblastoma cell lines induced to differentiation with all-trans retinoic acid. Mpped2 codes for a new metallophosphodiesterase protein, the expression of which inhibits cell proliferation and soft agar colony formation in SH -SY5Y cells. This inhibition is concomitant to an increased proportion of the cells in G0/G1 phase and enhanced caspase 3 activation, effects not seen for the other phosphodiesterases. A Mpped2-null mutation (H67R) abrogates these functions, which indicates that the biochemical activity of Mpped2 is advantageous for cancer suppression. Expression analyses in the "Los Angeles" and "Essen" neuroblastoma gene-array data sets show that increased expression of Mpped2 is associated with good patient prognosis according to Kaplan-Meier analyses. Tumorigenic assays in mice show that overexpression of Mpped2 improves survival rate, substantially impairs tumor growth and induces neuronal differentiation. Altogether, these data show that Mpped2 expression impairs neuroblastoma tumorigenesis, and they establish a basis for future therapeutic applications.

Gallbladder Polyps in Primary Sclerosing Cholangitis: Indication for Early Intervention

Analysis of Human Milk to Assess Exposure to PAHs, PCBs and Organochlorine Pesticides in the Vicinity Mediterranean City Mersin, Turkey

Assessment of human exposure to environmental persistent organic pollutants such as organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins/furans (PCDD/Fs) and polycyclic aromatic hydrocarbons (PAHs) based on the levels in human breast milk provides a reasonable tool not only to assess the contaminant burden in mothers but also to assess potential exposure of breast-fed neonates. There are limited data on levels of PCBs and OCPs in humans but no previous reports from Turkey on chemically determined levels PAHs in human milk. The aim of this study was to report the levels and accumulation profiles of OCPs, PCBs and PAHs in 47 breast milk samples obtained from a Mediterranean city, Mersin. High resolution analyses were performed by a gas chromatography coupled with mass spectrometer (GC-MS). Dichlorodiphenyldichloroethane (4.4'-DDE) was the dominant pollutant. Beta-hexachlorocyclohexane (ß-HCH), dichlorodiphenyltrichloroethane (4.4'-DDT), dieldrin, hexachlorobenzene, oxy-chlordane, cis-heptachlorepoxide were the other main OCPs detected. Mean levels of ΣPCB congeners and WHO(PCB)-TEQ were 9.94 and 0.001 ng/g lipid, respectively. PCB 153 showed the highest concentration (3.37 ng/g lipid), followed by PCB 138 and 180. For the dioxin-like PCBs, PCB 118 was the dominant (0.97 ng/g lipid). Naphthalene, phenanthrene, pyrene and fluoranthene were the major PAHs among the 16 PAHs detected. The estimated daily intakes of DDTs, PCBs, HCHs and HCB were not exceeded the tolerable daily intake (TDI) proposed by the Health Canada Guideline. These results indicate that the neonates of Mersin city are exposed to persistent organic pollutants analyzed in this study. However, neonates born in Mersin province are less exposed than the ones born in other regions, considering OCP and PCB levels in breast milk.

Fluorescence in Situ Hybridization (FISH) Detection of Nitrite Reductase Transcripts (nirS MRNA) in Pseudomonas Stutzeri Biofilms Relative to a Microscale Oxygen Gradient

Microsensor measurements of oxygen were combined with mRNA-targeted fluorescence in situ hybridization (FISH) to relate the expression of nitrite reductase (nirS) to oxygen concentrations in artificial biofilms of the denitrifier Pseudomonas stutzeri. A distinct zone of nirS transcript-containing cells was detected at the oxic-anoxic transition zone, below an oxygen threshold concentration of 0.7-2.5μM, depending on incubation conditions. Although not a routine technique yet, the possibility of coupling microsensor and mRNA-targeted FISH analyses described here opens for studies addressing microenvironment, identity, and actual activity of microbes in stratified environments at single cell resolution.

Mercury Levels in Feed and Muscle of Farmed Tilapia

Fish consumption is considered the most important source of contaminant exposure for humans beings, and farmed fish can be exposed to contaminants via feed supply.

Definitive Radiotherapy Versus Postoperative Radiotherapy of Patients with Oro- and Hypopharyngeal Cancer: Impact of Prognostic Factors

Purpose. To compare the impact of prognostic factors of patients treated with definitive radio(chemo)therapy versus patients treated with surgery and postoperative radiotherapy for squamous cell carcinoma of the oro- and hypopharynx. Patients and Methods. 162 patients treated with definitive radiotherapy and 126 patients treated with postoperative radiotherapy were retrospectively analysed. The impact of the prognostic factors gender, age, total tumor volume (TTV), pre-radiotherapy hemoglobin level (Hb-level), tumor site, T- and N-classification, radiotherapy interruptions >5 days, radiotherapy versus simultaneous radiochemotherapy, R-status and time interval between surgery and radiotherapy were investigated. Results. The median follow-up time for the censored patients treated with definitive radio(chemo)therapy was 28.5 months and for postoperative radiotherapy 36.5 months. On univariate analysis, the TTV, Hb-level, and simultaneous radiochemotherapy had a significant impact on the survival of patients treated with definitive radio(chemo)therapy. For patients treated with postoperative radiotherapy, only the TTV showed a statistical trend for the survival (P = 0.13). On multivariate analysis, the TTV and simultaneous radiochemotherapy maintained their statistical significance for patients treated with definitive raditherapy, and the TTV, the statistical trend for patients treated with postoperative radiotherapy (P = 0.19). Conclusions. The TTV was the predominant prognostic factor for both, patients treated with definitive or postoperative radiotherapy.

Clinical Impact and Predictors of Carotid Artery In-stent Restenosis

To assess the incidence and clinical significance as well as predictors of in-stent restenosis (ISR) after carotid artery stenting (CAS) diagnosed with serial duplex sonography investigations. We analyzed 215 CAS procedures that had clinical and serial carotid duplex ultrasound investigations. The incidence of in-stent restenosis (ISR) and periprocedural as well as long-term clinical complications were recorded. The influence of an ISR on clinical complication was analyzed using Kaplan-Meier curves and clinical risk factors for the development of an ISR with multivariate logistic regression. During a median follow-up time of 33.4 months (interquartile range 15.3-53.7) an ISR of ≥70% was detected in 12 (6.1%) of 215 arteries (mean age of 68.1 ± 9.8 years, 71.6% male). The combined stroke and death rate during long-term follow-up was significantly higher in the group with an ISR [odds ratio (OR): 3.59, 95% confidence interval (CI): 1.50-8.59, p = 0.004]. After applying multivariate logistic regression analysis contralateral carotid occlusion (OR 10.11, 95% CI 2.06-49.63, p = 0.004), carotid endarterectomy (CEA) restenosis (OR 8.87, 95% CI 1.68-46.84, p = 0.010) and postprocedural carotid duplex ultrasound with a PSV ≥120 cm/s (OR 6.33, 95% CI 1.27-31.44, p = 0.024) were independent predictors of ISR. ISR after CAS during long-term follow-up is associated with a higher proportion of clinical complications. A close follow-up is suggested especially in those patients with the aforementioned independent predictors of an ISR. Against the background of a lacking established treatment of ISR, these findings should be taken into account when offering CAS as a treatment alternative to CEA.

Neuroprotective Effects of a Taurine-containing Irrigation Solution for Vitrectomy

During pars plana vitrectomy, the retina is exposed to several iatrogenic risk factors, including excitotoxicity. A taurine-containing irrigation solution for pars plana vitrectomy (PURI PROTECT) has been developed and is claimed to have neuroprotective properties.

Spatial Distribution and Temporal Trend in Concentration of Carbofuran, Diazinon and Chlorpyrifos Ethyl Residues in Sediment and Water in Lake Naivasha, Kenya

Chlorpyrifos ethyl was found to be widely distributed in water and sediment in Lake Naivasha. Higher levels were reported in sediment (11.2-30.0 ng g(-1) dry weight (dw) in wet season than in dry season (4.7-17.4 ng g(-1) dw). The mean concentration of chlorpyrifos ethyl in water in wet season ranged between 8.8 and 26.6 μg L(-1) and decreased to between below detection limit to 14.0 μg L(-1) in dry season. On average, higher concentrations of chlorpyrifos ethyl were observed in sediment than water samples. Statistical analysis revealed a significant difference in concentration between the seasons, and a significant interaction between seasons and mean concentrations at p ≤ 0.05. However, levels of diazinon and carbofuran were below the detection limit in all the samples analyzed. Notably, levels of chlorpyrifos ethyl were higher than the maximum allowable limits (0.1 μg L(-1)) recommended by European Union for drinking water and general water quality criterion for protection of freshwater water organisms (0.083 μg L(-1)).

β-catenin is a Central Mediator of Pro-fibrotic Wnt Signaling in Systemic Sclerosis

Pathologic fibroblast activation drives fibrosis of the skin and internal organs in patients with systemic sclerosis (SSc). β-catenin is an integral part of adherens junctions and a central component of canonical Wnt signaling. Here, the authors addressed the role of β-catenin in fibroblasts for the development of SSc dermal fibrosis.

[The Hybrid Operating Room. Home of High-end Intraoperative Imaging]

A hybrid operating room must serve the medical needs of different highly specialized disciplines. It integrates interventional techniques for cardiovascular procedures and allows operations in the field of orthopaedic surgery, neurosurgery and maxillofacial surgery. The integration of all steps such as planning, documentation and the procedure itself saves time and precious resources. The best available imaging devices and user interfaces reduce the need for extensive personnel in the OR and facilitate new minimally invasive procedures. The immediate possibility of postoperative control images in CT-like quality enables the surgeon to react to problems during the same procedure without the need for later revision.

Purifying Selection and Molecular Adaptation in the Genome of Verminephrobacter, the Heritable Symbiotic Bacteria of Earthworms

While genomic erosion is common among intracellular symbionts, patterns of genome evolution in heritable extracellular endosymbionts remain elusive. We study vertically transmitted extracellular endosymbionts (Verminephrobacter, Betaproteobacteria) that form a beneficial, species-specific, and evolutionarily old (60-130 Myr) association with earthworms. We assembled a draft genome of Verminephrobacter aporrectodeae and compared it with the genomes of Verminephrobacter eiseniae and two nonsymbiotic close relatives (Acidovorax). Similar to V. eiseniae, the V. aporrectodeae genome was not markedly reduced in size and showed no A-T bias. We characterized the strength of purifying selection (ω = dN/dS) and codon usage bias in 876 orthologous genes. Symbiont genomes exhibited strong purifying selection (ω = 0.09 ± 0.07), although transition to symbiosis entailed relaxation of purifying selection as evidenced by 50% higher ω values and less codon usage bias in symbiont compared with reference genomes. Relaxation was not evenly distributed among functional gene categories but was overrepresented in genes involved in signal transduction and cell envelope biogenesis. The same gene categories also harbored instances of positive selection in the Verminephrobacter clade. In total, positive selection was detected in 89 genes, including also genes involved in DNA metabolism, tRNA modification, and TonB-dependent iron uptake, potentially highlighting functions important in symbiosis. Our results suggest that the transition to symbiosis was accompanied by molecular adaptation, while purifying selection was only moderately relaxed, despite the evolutionary age and stability of the host association. We hypothesize that biparental transmission of symbionts and rare genetic mixing during transmission can prevent genome erosion in heritable symbionts.

Advances and Innovations in Computer-assisted Head and Neck Oncologic Surgery

ABSTRACT: Over the past years, computer-assisted surgery has gained more importance in craniomaxillofacial surgery, especially in primary and secondary treatment of head and neck malignancies. The basis for oncologic treatment of the head and neck region requires detailed planning using computed tomography, cone-beam computed tomography, or magnetic resonance imaging in combination with computer-assisted, infrared-based navigation system. These techniques allow a preplanned image-guided path to the tumor region for taking biopsies, resection, or reconstruction. The aim of this work was to show the advances and technical benefits for tumor surgery in a daily clinical routine from the view of the craniomaxillofacial surgeon. The target of our working group was to develop and clinically evaluate a novel three-dimensional planning and navigation software solution for treatment of craniofacial tumors.This work was carried out on 5 categories for oncologic surgical procedures in which computer-assisted surgery was applied from 2005 to 2011: preplanned trajectorial-guided tumor biopsy, intraoperative image-controlled tumor resection, tumor mapping, reconstruction after tumor surgery (true to original), and oral rehabilitation (backward planning). Successful preoperative planning, import of image data suitable for navigation, and intraoperative precise infrared-based navigation were obtained for all 5 categories without any complications.Image-guided navigation technique for head and neck oncologic surgery provides a precise, safe surgical method with real-time excellent anatomic orientation. Regarding the advantages of computer-assisted surgery, this technique will play a major part in craniofacial reconstructive surgery and will address widespread general methodologic solutions that are of great interest in multidisciplinary oncologic treatment.

[2003 Nursing Regulation and the Sequelae: Plea for Reform: a Short Guest Performance on the Unit]

Comparison of Outcomes in Children with Hearing Aids and Cochlear Implants

The purpose of this study was to document the performance of a group of children with moderately severe to severe hearing loss who use hearing aids on a range of speech recognition, speech-language, and literacy measures and to compare these results to children with severe to profound hearing loss, who have learned language through cochlear implants.

5-Fluorouracil Accumulation in Green Microalgae and Its Biogenetic Transfer into Ciliate Protozoan

The study has demonstrated that anticancer drug 5-fluorouracil causes acute toxicity and interferes with the growth of green microalgae, Scenedesmus vacuolatus. It accumulates in microalgae biomass with bioaccumulation factor of 1.84 × 10(4) and further integrates into the DNA and RNA of microalgae. In addition, the labelled microalgae genome is transferred into protozoan Tetrahymena pyriformis on feeding and is retained in the food vacuoles of predator organisms. This biotransfer of labelled 5-fluorouracil via genomic material was evaluated using radioactivity in Tetrahymena cell pellets though radioactivity did not detect anticancer drug in the genome of the predator organism.

The Lateral Thoracic Nerve and the Cutaneous Maximus Muscle-A Novel in Vivo Model System for Nerve Degeneration and Regeneration Studies

We report a novel in vivo mouse model system to study regeneration of injured motor nerve and spatiotemporal pattern of denervation in experimental nerve diseases. The lateral thoracic nerve (LTN), as a pure motor nerve, innervates the cutaneous maximus muscle (CMM) by some of the shortest and the longest motor nerve fibers in the mouse body. Its branches and nerve terminals can be imaged in whole mount preparations. Here we describe the branching pattern of the LTN and its innervation of the CMM, and characterize degeneration and regeneration over time after a LTN crush by morphological and electrophysiological analyses. We demonstrate the utility of this model in a well-established neurotoxicity paradigm and in a genetic disease model of the peripheral neuropathy. Furthermore, this system enables punch biopsies that allow repeated and multi-location examinations for LTN regeneration and CMM reinnervation over time. The presence of the LTN and the CMM in a variety of species and its easy accessibility suggests that this in vivo model system offers considerable promise for future nerve degeneration and regeneration research.

The Influence of Post-growth Annealing on the Optical Properties of InAs Quantum Dot Chains Grown on Pre-patterned GaAs(100)

We report on the effect of post-growth thermal annealing of [011]- ,[011(-)]-, and [010]-oriented quantum dot chains grown by molecular beam epitaxy on GaAs(100) substrates patterned by UV-nanoimprint lithography. We show that the quantum dot chains experience a blueshift of the photoluminescence energy, spectral narrowing, and a reduction of the intersubband energy separation during annealing. The photoluminescence blueshift is more rapid for the quantum dot chains than for self-assembled quantum dots that were used as a reference. Furthermore, we studied polarization resolved photoluminescence and observed that annealing reduces the intrinsic optical anisotropy of the quantum dot chains and the self-assembled quantum dots.

Analysis of Optical Side-effects of Fs-laser Therapy in Human Presbyopic Lens Simulated with Modified Contact Lenses

In a single-blinded study, optical side-effects of a potential femtosecond (fs)-laser therapy in presbyopic human lenses were tested. Simulation of this therapy was carried out by applying fs-laser patterns into standard contact lenses (CL).

Continuous Regional Myocardial Blood Flow Measurement: Validation of a Near-infrared Laser Doppler Device in a Porcine Model

RMBF measurement is a major concern in various clinical and experimental settings, but no validated device for RMBF is currently available. Methods:  An LVP-triggered laser Doppler to measure RMBF was validated by simultaneous fluorescent MS RMBF in a porcine LAD flow reduction model (n = 10 pigs). The laser probe was positioned on the left ventricle's anterior wall. LAD blood flow reduction was achieved by a shaft-driven occluder positioned proximal to the transit-time flow meter measuring coronary blood flow. RMBF was measured at baseline; after the reduction of LAD blood flow to 70% and 30% of baseline; at 20 and 120 minutes of reperfusion; and, finally, 15 minutes after LAD occlusion.

Cellular Pharmacology of Protein Kinase Mζ (PKMζ) Contrasts with Its in Vitro Profile: Implications for PKMζ As a Mediator of Memory

A number of recent studies have used pharmacological inhibitors to establish a role for protein kinase Mζ (PKMζ) in synaptic plasticity and memory. These studies use zeta inhibitory peptide (ZIP) and chelerythrine as inhibitors of PKMζ to block long term potentiation and memory; staurosporine is used as a negative control to show that a nonspecific kinase inhibitor does not block long term potentiation and memory. Here, we show that neither ZIP nor chelerythrine inhibits PKMζ in cultured cells or brain slices. In contrast, staurosporine does block PKMζ activity in cells and brain slices by inhibiting its upstream phosphoinositide-dependent kinase-1. These studies demonstrate that the effectiveness of drugs against purified PKMζ may not be indicative of their specificity in the more complex environment of the cell and suggest that PKMζ is unlikely to be the mediator of synaptic plasticity or memory.

Succession of Deferribacteres and Epsilonproteobacteria Through a Nitrate-treated High-temperature Oil Production Facility

Members of Epsilonproteobacteria and Deferribacteres have been implied in nitrate-induced souring control in high-temperature oil production facilities. Here we report on their diversity and abundance in the injection and production part of a nitrate-treated, off-shore oil facility (Halfdan, Denmark) and aimed to assess their potential in souring control. Nitrate addition to deoxygenated seawater shifted the low-biomass seawater community dominated by Gammaproteobacteria closely affiliated with the genus Colwellia to a high-biomass community with significantly higher species richness. Epsilonproteobacteria accounted for less than 1% of the total bacterial community in the nitrate-amended injection water and were most likely outcompeted by putative nitrate-reducing, methylotrophic Gammaproteobacteria of the genus Methylophaga. Reservoir passage and recovery of the oil resulted in a significant change in the bacterial community. Members of the thermophilic Deferribacteres were the second major fraction of the bacterial community in the production water (~30% of the total bacterial community). They were not found in the injection water and were therefore assumed to be indigenous to the reservoir. Additional diversity analysis and targeted quantification of periplasmic nitrate reductase (napA) genes indicated that most resident Deferribacteres possessed the functional potential to contribute to nitrate reduction in the system. In sum, the dominance of nitrate-reducing Deferribacteres and the low relative abundance of Epsilonproteobacteria throughout the production facility suggested that the Deferribacteres play a major role in nitrate-induced souring control at high temperatures.

The Mammalian Synaptonemal Complex: Protein Components, Assembly and Role in Meiotic Recombination

The synaptonemal complex (SC) is a proteinaceous structure of chromosome bivalents whose assembly is indispensable for the successful progression of the first meiotic division of sexually reproducing organisms. In this mini-review we will focus on recent progress dealing with the composition and assembly of the mammalian SC. These advances mainly resulted from the systematic use of knockout mice for all known mammalian SC proteins as well as from protein polymerization studies performed in heterologous systems.

Hyperpolarization-activated Cation Current Ih of Dentate Gyrus Granule Cells is Upregulated in Human and Rat Temporal Lobe Epilepsy

The hyperpolarization-activated cation current I(h) is an important regulator of neuronal excitability and may contribute to the properties of the dentate gyrus granule (DGG) cells, which constitute the input site of the canonical hippocampal circuit. Here, we investigated changes in I(h) in DGG cells in human temporal lobe epilepsy (TLE) and the rat pilocarpine model of TLE using the patch-clamp technique. Messenger-RNA (mRNA) expression of I(h)-conducting HCN1, 2 and 4 isoforms was determined using semi-quantitative in-situ hybridization. I(h) density was ∼1.8-fold greater in DGG cells of TLE patients with Ammon's horn sclerosis (AHS) as compared to patients without AHS. The magnitude of somatodendritic I(h) was enhanced also in DGG cells in epileptic rats, most robustly during the latent phase after status epilepticus and prior to the occurrence of spontaneous epileptic seizures. During the chronic phase, I(h) was increased ∼1.7-fold. This increase of I(h) was paralleled by an increase in HCN1 and HCN4 mRNA expression, whereas HCN2 expression was unchanged. Our data demonstrate an epilepsy-associated upregulation of I(h) likely due to increased HCN1 and HCN4 expression, which indicate plasticity of I(h) during epileptogenesis and which may contribute to a compensatory decrease in neuronal excitability of DGG cells.

Targeting NADPH Oxidases in Vascular Pharmacology

Oxidative stress is a molecular dysregulation in reactive oxygen species (ROS) metabolism, which plays a key role in the pathogenesis of atherosclerosis, vascular inflammation and endothelial dysfunction. It is characterized by a loss of nitric oxide (NO) bioavailability. Large clinical trials such as HOPE and HPS have not shown a clinical benefit of antioxidant vitamin C or vitamin E treatment, putting into question the role of oxidative stress in cardiovascular disease. A change in the understanding of the molecular nature of oxidative stress has been driven by the results of these trials. Oxidative stress is no longer perceived as a simple imbalance between the production and scavenging of ROS, but as a dysfunction of enzymes involved in ROS production. NADPH oxidases are at the center of these events, underlying the dysfunction of other oxidases including eNOS uncoupling, xanthine oxidase and mitochondrial dysfunction. Thus NADPH oxidases are important therapeutic targets. Indeed, HMG-CoA reductase inhibitors (statins) as well as drugs interfering with the renin-angiotensin-aldosterone system inhibit NADPH oxidase activation and expression. Angiotensin-converting enzyme (ACE) inhibitors, AT1 receptor antagonists (sartans) and aliskiren, as well as spironolactone or eplerenone, have been discussed. Molecular aspects of NADPH oxidase regulation must be considered, while thinking about novel pharmacological targeting of this family of enzymes consisting of several homologs Nox1, Nox2, Nox3, Nox4 and Nox5 in humans. In order to properly design trials of antioxidant therapies, we must develop reliable techniques for the assessment of local and systemic oxidative stress. Classical antioxidants could be combined with novel oxidase inhibitors. In this review, we discuss NADPH oxidase inhibitors such as VAS2870, VAS3947, GK-136901, S17834 or plumbagin. Therefore, our efforts must focus on generating small molecular weight inhibitors of NADPH oxidases, allowing the selective inhibition of dysfunctional NADPH oxidase homologs. This appears to be the most reasonable approach, potentially much more efficient than non-selective scavenging of all ROS by the administration of antioxidants.

Luciferase-based Assay for Adenosine: Application to S-adenosyl-L-homocysteine Hydrolase

S-Adenosyl-L-homocysteine hydrolase (SAHH) catalyzes the reversible conversion of S-adenosyl-L-homocysteine (SAH) to adenosine (ADO) and L-homocysteine, promoting methyltransferase activity by relief of SAH inhibition. SAH catabolism is linked to S-adenosylmethionine metabolism, and the development of SAHH inhibitors is of interest for new therapeutics with anticancer or cholesterol-lowering effects. We have developed a continuous enzymatic assay for adenosine that facilitates high-throughput analysis of SAHH. This luciferase-based assay is 4000-fold more sensitive than former detection methods and is well suited for continuous monitoring of ADO formation in a 96-well-plate format. The high-affinity adenosine kinase from Anopheles gambiae efficiently converts adenosine to adenosine monophosphate (AMP) in the presence of guanosine triphosphate. AMP is converted to adenosine triphosphate and coupled to firefly luciferase. With this procedure, kinetic parameters (K(m), k(cat)) for SAHH were obtained, in good agreement with literature values. Assay characteristics include sustained light output combined with ultrasensitive detection (10(-7) unit of SAHH). The assay is documented with the characterization of slow-onset inhibition for inhibitors of the hydrolase. Application of this assay may facilitate the development of SAHH inhibitors and provide an ultrasensitive detection for the formation of adenosine from other biological reactions.

Improved Anatomic Visualization of a Glomus Caroticum Tumour Within the Carotic Bifurcation with Combined 68Ga-DOTATATE PET/MRI

Familial Occurrence of the VATER/VACTERL Association

The acronym VATER/VACTERL association is used to describe the non-random co-occurrence of vertebral defects (V), anorectal malformations (A), cardiac defects (C), tracheo-esophageal fistula with or without esophageal atresia (TE), renal malformations (R), and limb defects (L). We report a familial case of VATER/VACTERL association in which both the index case and her maternal uncle displayed four major component features of the disorder. A systematic literature search identified 12 previously described familial cases. However, on comparison, both members fulfilled the diagnostic criteria for VATER/VACTERL association only in one instance, and ours is the second such report. Although, a SNP array-based analysis identified no causal genomic alteration, the findings in the present family suggest that genetic factors are implicated in the development of the disorder.

FOXP3+ Regulatory T Cells in Autoimmune Hepatitis Are Fully Functional and Not Reduced in Frequency

The pathogenesis of autoimmune hepatitis (AIH) is not understood, but it was suggested that AIH may be related to a numerical or functional impairment of CD4+CD25+FOXP3+ regulatory T cells (Treg), which are important mediators of immune tolerance to self-antigens. However, the role of Treg in AIH is not clear, since earlier studies reporting Treg impairment had used only CD25 as marker that cannot unambiguously distinguish Treg from activated effector T cells.

Sensitivity to Detect Change and the Correlation of Clinical Factors with the Hamilton Depression Rating Scale and the Beck Depression Inventory in Depressed Inpatients

Discrepancies between scores on the Hamilton Depression Rating Scale (HAMD) and the Beck Depression Inventory (BDI), as well as differences regarding their sensitivity to detect change, have been reported. This study investigates discrepancies and their potential prediction on the basis of demographic, personality, and clinical factors in depressed inpatients and analyzes the sensitivity to change. The HAMD and the BDI were administered to 105 inpatients with major depressive disorder randomized to 5 weeks of either interpersonal psychotherapy or clinical management. Personality was assessed with the NEO Five-Factor Inventory. Low extraversion and high neuroticism were associated with relatively higher endorsement of depressive symptoms on the BDI compared with the HAMD. The HAMD presented a greater reduction of symptom scores than the BDI. Patients with high BDI scores, high HAMD scores or both revealed the greatest change, possibly due to a statistical effect of regression to the mean. Restricted by sample size, analyses were not differentiated by treatment condition. Regression to the mean cannot be tested directly, but it might be considered as a possible explanation. The HAMD and the BDI should be regarded as two complementary rather than redundant or competing instruments as the discrepancy is associated with personality characteristics. Attributing large effect sizes solely to effective treatment and a sensitive measure may be misleading.

Evaluating Retinal Toxicity of a New Heavy Intraocular Dye, Using a Model of Perfused and Isolated Retinal Cultures of Bovine and Human Origin

Dyes such as brilliant blue (BBG) are used during vitreoretinal surgery to visualize anatomical structures. By adding deuterium oxide (D2O), surgeons have tried to create a dye mixture heavier than water to facilitate staining of the inner limiting membrane (ILM) without prior fluid-air exchange. This study investigated the effect of 0.4 ml BBG (Fluoron, Ulm, Germany) mixed with 0.13 ml/ml D2O and D2O on retinal function of a pseudo in vivo model using bovine and human whole mount cultures.

Phenotypic Changes to the Endogenous Antigen-specific CD8+ T Cell Response Correlates with the Development and Resolution of Allergic Airway Disease

The role of CD8(+) T cells in the pathogenesis of asthma remains controversial, as both pro- and anti-inflammatory functions have been suggested. This study was designed to examine the endogenous CD8(+) T cell response in a biphasic ovalbumin (OVA)-induced model of allergic airway disease (AAD) and its subsequent resolution with the development of local inhalational tolerance (LIT). We observed increases in OVA-specific CD8(+) T cell numbers in the local lung compartments (bronchoalveolar lavage, lung tissue, hilar lymph node) at AAD and LIT; systemic compartments (spleen, inguinal lymph node) displayed no such increases in CD8(+) T cell numbers. OVA-specific CD8(+) T cells appeared to exhibit plasticity both phenotypically and functionally. They possessed pro-inflammatory characteristics at AAD, with high phenotypic expression of CD11a and increased functional expression of granzyme B and interferon-γ. In contrast, at LIT they showed increased phenotypic expression of the inhibitory marker NKG2A and functionally did not produce granzyme B or interferon-γ. In addition, in a discontinuous model the OVA-specific CD8(+) T cells could be recalled on re-exposure to OVA, demonstrating memory. Finally, confocal microscopy results showed that OVA-specific CD8(+) T cells at AAD are associated with B cell aggregates in lung tissue. These B cell aggregates resembled tertiary ectopic lymphoid tissue and may thus provide a local environment for the salient cellular interactions that contribute to the development of LIT.

Highly Adaptable Two-dimensional Metal-organic Coordination Networks on Metal Surfaces

The formation of extended two-dimensional metal-organic coordination networks (2D-MOCNs) showing high adaptability to surface step edges and structural defects is revealed by scanning tunneling microscopy. Rod-like 4,4'-di-(1,4-buta-1,3-diynyl)-benzoic acid (BDBA) and iron atoms assemble into extended 2D-MOCNs on Au(111) and Ag(100) surfaces. Independent from the chosen substrate and its surface symmetry the MOCN grows continuously over multiple surface terraces through mutual in-phase structure adaptation of network domains at step edges as well as on terraces. The adaptability of the MOCNs is mainly ascribed to the high degree of conformational flexibility of the butadiynyl functionality of the ligand. Despite their flexibility, the MOCNs exhibit considerable robustness against annealing at high temperatures. The findings show that mesoscale self-assembled functional architectures with a high degree of substrate error tolerance can be realized with metal coordination networks.

Primate Genome Gain and Loss: a Bone Dysplasia, Muscular Dystrophy, and Bone Cancer Syndrome Resulting from Mutated Retroviral-derived MTAP Transcripts

Diaphyseal medullary stenosis with malignant fibrous histiocytoma (DMS-MFH) is an autosomal-dominant syndrome characterized by bone dysplasia, myopathy, and bone cancer. We previously mapped the DMS-MFH tumor-suppressing-gene locus to chromosomal region 9p21-22 but failed to identify mutations in known genes in this region. We now demonstrate that DMS-MFH results from mutations in the most proximal of three previously uncharacterized terminal exons of the gene encoding methylthioadenosine phosphorylase, MTAP. Intriguingly, two of these MTAP exons arose from early and independent retroviral-integration events in primate genomes at least 40 million years ago, and since then, their genomic integration has gained a functional role. MTAP is a ubiquitously expressed homotrimeric-subunit enzyme critical to polyamine metabolism and adenine and methionine salvage pathways and was believed to be encoded as a single transcript from the eight previously described exons. Six distinct retroviral-sequence-containing MTAP isoforms, each of which can physically interact with archetype MTAP, have been identified. The disease-causing mutations occur within one of these retroviral-derived exons and result in exon skipping and dysregulated alternative splicing of all MTAP isoforms. Our results identify a gene involved in the development of bone sarcoma, provide evidence of the primate-specific evolution of certain parts of an existing gene, and demonstrate that mutations in parts of this gene can result in human disease despite its relatively recent origin.

The Facilitating Effect of Clinical Hypnosis on Motor Imagery: an FMRI Study

Hypnosis is increasingly being employed in therapy of neurologically impaired patients. In fact, reports from neuropsychological practice point out that neurological patients with a loss of motor abilities achieve successful rehabilitation by means of motor imagery during hypnosis. This approach was shown to be effective even if the patients' ability to imagine movements was impaired or lost. The underlying mechanisms of "how" and "where" hypnosis affects the brain, however, are largely unknown. To identify the brain areas involved in motor imagery under hypnosis, we conducted an fMRI study in which we required healthy human subjects either to imagine or to execute repetitive finger movements during a hypnotic trance. We observed fMRI-signal increases exclusively related to hypnosis in the left superior frontal cortex, the left anterior cingulate gyrus and left thalamus. While the superior frontal cortex and the anterior cingulate were active related more to movement performance than to imagery, the thalamus was activated only during motor imagery. These areas represent central nodes of the salience network linking primary and higher motor areas. Therefore, our data substantiate the notion that hypnosis enhances motor imagery.

[Supported Own Initiative of Rehabilitation Patients ("new Credo") - a Successful Model?]

In recent years, rehabilitation aftercare has become an important topic in rehabilitation research and practice. This development was initiated by the lack of long-term effects after rehabilitation treatment and by the deficits identified in several reviews of aftercare recommendations and of actual practice of aftercare. Against this background, a new aftercare strategy was developed, the "New Credo", which focused aftercare from the beginning of rehabilitation treatment. A substantial element of the concept is the resumption and maintenance of physical activity in everyday life of the rehabilitation patient. The goal of the present study was the evaluation of this concept in a sample of patients with chronic back pain.The "New Credo" was evaluated in a multi-centre, prospective controlled longitudinal study with 3 points of measurement (before and after rehab as well as after 12 months). 6 rehabilitation clinics in Schleswig-Holstein were included in the intervention arm (IG, 3 clinics) and in the control arm (CG, 3 clinics), participants were rehabilitants with chronic back pain. The clinics in the intervention arm implemented "the New Credo" in accordance with their local conditions. During the study period they were provided with a person who was responsible for managing aftercare activities for patients. Participants in the CG received standard rehabilitation treatment according to general guidelines. Restriction of social participation, disability days and restriction in functional capacity in everyday life were defined as primary outcomes. Complete data were available for 166 rehabilitants in the IG and 368 in the CG.At the end of rehabilitation rehabilitants in the IG reported significantly higher use of treatment offerings and more intensive preparation for the time after rehabilitation treatment than rehabilitants in the CG. Rehabilitants in the IG found recommendations for the time after treatment significantly better than those in the CG. As expected, both rehabilitants in the IG and in the CG showed similar outcomes directly after inpatient treatment. With respect to long-term effects 12 months later, significant differences with moderate effect sizes were observed in 2 of the 3 primary outcomes (function capacity and social participation) in favour of the IG. With respect to secondary outcomes, rehabilitants in the IG showed significantly better long-term results than those in the CG.Our results show that the "New Credo" can successfully be implemented into rehabilitation practice. The "New Credo" was positively rated by clinicians and rehabilitants. Rehabilitants in the IG found offerings of supported aftercare very helpful. The intensity of actual use of aftercare by rehabilitants as well as the emphasis laid on aftercare by clinicians indicate that treatment was focused on aftercare over the whole time of rehabilitation and that the rehabilitants transferred these aspects into everyday life. For the rehabilitants in the IG higher long-term effects could be observed compared to those in the CG. This suggests an improved sustainability of rehabilitation effects and a successful transfer of acquired knowledge and self-efficacy in everyday life.

Rationally Designing Safer Anilines: the Challenging Case of 4-aminobiphenyls

We describe how we have been able to design 4-aminobiphenyls that are nonmutagenic (inactive in the Ames test). No such 4-aminobiphenyls were known to us, but insights provided by quantum mechanical calculations have permitted us to design and synthesize some examples. Importantly, the quantum mechanical calculations could be combined with predictions of other properties of the compounds that contained the 4-aminobiphenyls so that these remained druglike. Having found compounds that are not active, the calculations can provide insight into which factors (electronic and conformational in this case) are important. The calculations provided SAR-like information that was able guide the design of further examples of 4-aminobiphenyls that are not active in the Ames test.

Large Array of Single, Site-controlled InAs Quantum Dots Fabricated by UV-nanoimprint Lithography and Molecular Beam Epitaxy

We present the growth of single, site-controlled InAs quantum dots on GaAs templates using UV-nanoimprint lithography and molecular beam epitaxy. A large quantum dot array with a period of 1.5 µm was achieved. Single quantum dots were studied by steady-state and time-resolved micro-photoluminescence experiments. We obtained single exciton emission with a linewidth of 45 µeV. In time-resolved experiments, we observed decay times of about 670 ps. Our results underline the potential of nanoimprint lithography and molecular beam epitaxy to create large-scale, single quantum dot arrays.

MYCN and ALKF1174L Are Sufficient to Drive Neuroblastoma Development from Neural Crest Progenitor Cells

Neuroblastoma is an embryonal tumor with a heterogeneous clinical course. The tumor is presumed to be derived from the neural crest, but the cells of origin remain to be determined. To date, few recurrent genetic changes contributing to neuroblastoma formation, such as amplification of the MYCN oncogene and activating mutations of the ALK oncogene, have been identified. The possibility to model neuroblastoma in mice allows investigation of the cell of origin hypothesis in further detail. Here we present the evidence that murine neural crest progenitor cells can give rise to neuroblastoma upon transformation with MYCN or ALK(F1174L). For this purpose we used JoMa1, a multipotent neural crest progenitor cell line, which is kept in a viable and undifferentiated state by a tamoxifen-activated c-Myc transgene (c-MycER(T)). Expression of MYCN or ALK(F1174L), one of the oncogenic ALK variants identified in primary neuroblastomas, enabled these cells to grow independently of c-MycER(T) activity in vitro and caused formation of neuroblastoma-like tumors in vivo in contrast to parental JoMa1 cells and JoMa1 cells-expressing TrkA or GFP. Tumorigenicity was enhanced upon serial transplantation of tumor-derived cells, and tumor cells remained susceptible to the MYC-inhibitor, NBT-272, indicating that cell growth depended on functional MYCN. Our findings support neural crest progenitor cells as the precursor cells of neuroblastoma, and indicate that neuroblastomas arise as their malignant progeny.Oncogene advance online publication, 9 April 2012; doi:10.1038/onc.2012.106.

Smac Mimetic LBW242 Sensitizes XIAP-overexpressing Neuroblastoma Cells for TNF-α-independent Apoptosis

Despite intensive treatment regimens, high-risk and late-stage neuroblastoma tends to have a poor survival outcome. Overexpression of the apoptotic regulator, X-linked inhibitor of apoptosis protein (XIAP), has been associated with chemotherapy resistance in several cancers including neuroblastoma. Here, we report preclinical evidence that XIAP offers an effective therapeutic target in neuroblastoma. Human and murine neuroblastoma cells were treated with the Smac mimetic LBW242 alone or in combination with cytotoxic drugs used clinically to treat neuroblastoma. Expression of XIAP protein, but not mRNA, was highly increased in neuroblastoma cells compared to healthy adrenal gland tissue, consistent with a posttranscriptional regulation of XIAP expression. Treatment with LBW242 sensitized human and murine neuroblastoma cells to chemotherapy-induced apoptosis, which was mediated by activation of both the intrinsic and extrinsic apoptosis pathways. Although Smac mimetics have been reported to stimulate TNF-α-induced apoptosis by degradation of cellular IAP (cIAP)-1/2, we found that LBW242-mediated sensitization in neuroblastoma cells occurred in a TNF-α-independent manner, despite induction of cIAP-1/2 degradation and TNF-α expression. Together, our findings show that XIAP targeting sensitizes neuroblastoma to chemotherapy-induced apoptosis, suggesting a novel therapeutic approach to treat this childhood malignancy.

Shell Biofilm Nitrification and Gut Denitrification Contribute to Emission of Nitrous Oxide by the Invasive Freshwater Mussel Dreissena Polymorpha (zebra Mussel)

Nitrification in shell biofilms and denitrification in the gut of the animal accounted for N(2)O emission by Dreissena polymorpha (Bivalvia), as shown by gas chromatography and gene expression analysis. The mussel's ammonium excretion was sufficient to sustain N(2)O production and thus potentially uncouples invertebrate N(2)O production from environmental N concentrations.

Transition States of Native and Drug-resistant HIV-1 Protease Are the Same

HIV-1 protease is an important target for the treatment of HIV/AIDS. However, drug resistance is a persistent problem and new inhibitors are needed. An approach toward understanding enzyme chemistry, the basis of drug resistance, and the design of powerful inhibitors is to establish the structure of enzymatic transition states. Enzymatic transition structures can be established by matching experimental kinetic isotope effects (KIEs) with theoretical predictions. However, the HIV-1 protease transition state has not been previously resolved using these methods. We have measured primary (14)C and (15)N KIEs and secondary (3)H and (18)O KIEs for native and multidrug-resistant HIV-1 protease (I84V). We observed (14)C KIEs ((14)V/K) of 1.029 ± 0.003 and 1.025 ± 0.005, (15)N KIEs ((15)V/K) of 0.987 ± 0.004 and 0.989 ± 0.003, (18)O KIEs ((18)V/K) of 0.999 ± 0.003 and 0.993 ± 0.003, and (3)H KIEs ((3)V/K) KIEs of 0.968 ± 0.001 and 0.976 ± 0.001 for the native and I84V enzyme, respectively. The chemical reaction involves nucleophilic water attack at the carbonyl carbon, proton transfer to the amide nitrogen leaving group, and C-N bond cleavage. A transition structure consistent with the KIE values involves proton transfer from the active site Asp-125 (1.32 Å) with partial hydrogen bond formation to the accepting nitrogen (1.20 Å) and partial bond loss from the carbonyl carbon to the amide leaving group (1.52 Å). The KIEs measured for the native and I84V enzyme indicate nearly identical transition states, implying that a true transition-state analogue should be effective against both enzymes.

[The First 24 Hours: Acute Dyspnea]

Acute dyspnea is a common presentation in the emergency department. Immediate diagnostic strategy and efficient management is crucial. Therefore, a diagnostic work up consisting of a brief medical history, physical examination and technical investigations, including laboratory tests, is presented. Identification of the cardio-vascular, pulmonary or other etiology enables the initiation of adequate therapy. This is outlined in detail for three common entities.

An Unusual Cause of Jaundice

Haemophilia Prophylaxis: How Can We Justify the Costs?

Severe haemophilia results in increased mortality and poorer quality of life. Factor prophylaxis leads to a more normal life, but is very costly; most of the cost is due to the high cost of replacement factor. Despite its high cost, factor prophylaxis has been adopted throughout the developed world--even in different health care systems. We argue that there are at least five possible reasons why societies may value factor prophylaxis despite its cost: (i) it is directed towards an inherited disease, (ii) the treatment is largely directed towards children, (iii) the disease is rare and the overall cost to society is small, (iv) the treatment is preventative, and v) the high cost is largely the result of providing safe products. In an era of rising health care costs, there is a strong research agenda to establish the factors that determine the value of expensive therapies for rare diseases like haemophilia.

Comments on an Article by Kamalian Et Al

Association Between Chemical Pattern in Breast Milk and Congenital Cryptorchidism: Modelling of Complex Human Exposures

During the past four decades, there has been an increase in the incidence rate of male reproductive disorders in some, but not all, Western countries. The observed increase in the prevalence of male reproductive disorders is suspected to be ascribable to environmental factors as the increase has been too rapid to be explained by genetics alone. To study the association between complex chemical exposures of humans and congenital cryptorchidism, the most common malformation of the male genitalia, we measured 121 environmental chemicals with suspected or known endocrine disrupting properties in 130 breast milk samples from Danish and Finnish mothers. Half the newborns were healthy controls, whereas the other half was boys with congenital cryptorchidism. The measured chemicals included polychlorinated biphenyls (PCBs), polybrominated diphenyl-ethers, dioxins (OCDD/PCDFs), phthalates, polybrominated biphenyls and organochlorine pesticides. Computational analysis of the data was performed using logistic regression and three multivariate machine learning classifiers. Furthermore, we performed systems biology analysis to explore the chemical influence on a molecular level. After correction for multiple testing, exposure to nine chemicals was significantly different between the cases and controls in the Danish cohort, but not in the Finnish cohort. The multivariate analysis indicated that Danish samples exhibited a stronger correlation between chemical exposure patterns in breast milk and cryptorchidism than Finnish samples. Moreover, PCBs were indicated as having a protective effect within the Danish cohort, which was supported by molecular data recovered through systems biology. Our results lend further support to the hypothesis that the mixture of environmental chemicals may contribute to observed adverse trends in male reproductive health.

Extended Analysis of a Genome-wide Association Study in Primary Sclerosing Cholangitis Detects Multiple Novel Risk Loci

A limited number of genetic risk factors have been reported in primary sclerosing cholangitis (PSC). To discover further genetic susceptibility factors for PSC, we followed up on a second tier of single nucleotide polymorphisms (SNPs) from a genome-wide association study (GWAS).

Strategies for Responding to RAC Requests Electronically

Providers that would like to respond to complex RAC reviews electronically should consider three strategies: Invest in an EHR software package or a high-powered scanner that can quickly scan large amounts of paper. Implement an audit software platform that will allow providers to manage the entire audit process in one place. Use a CONNECT-compatible gateway capable of accessing the Nationwide Health Information Network (the network on which the electronic submission of medical documentation program runs).

Management of Bleeding Disorders in the Elderly

We are entering a new phase in the management of patients with bleeding disorders such as haemophilia. This is the result of the positive effects that disease management strategies have had on patient longevity over the last 10-15 years. A greater number of individuals are entering middle- to old-age and, as a result, we face a new era of having to manage haemophiliac patients at risk of, or suffering from, age-related diseases. We can clearly learn from the experiences of geriatricians who have made many advances in the management of chronic disorders such as cardiovascular diseases and osteoporosis. However, the hypocoagulable state brings challenges of its own and it is important that we communicate our experiences so that the shared information can help drive improved levels of care and better clinical outcomes. In this article we look at factors that have impacted the life expectancy of patients with haemophilia over the last few decades, and we also review some of the early literature relating to cardiovascular risk management and the treatment of osteoporosis.

[52-year-old Man with Pain of the Neck]

Falls Prevention for the Elderly

An ageing population, a growing prevalence of chronic diseases and limited financial resources for health care underpin the importance of prevention of disabling health disorders and care dependency in the elderly. A wide variety of measures is generally available for the prevention of falls and fall-related injuries. The spectrum ranges from diagnostic procedures for identifying individuals at risk of falling to complex interventions for the removal or reduction of identified risk factors. However, the clinical and economic effectiveness of the majority of recommended strategies for fall prevention is unclear. Against this background, the literature analyses in this HTA report aim to support decision-making for effective and efficient fall prevention.

Design of a Multi-signature Ensemble Classifier Predicting Neuroblastoma Patients' Outcome

Neuroblastoma is the most common pediatric solid tumor of the sympathetic nervous system. Development of improved predictive tools for patients stratification is a crucial requirement for neuroblastoma therapy. Several studies utilized gene expression-based signatures to stratify neuroblastoma patients and demonstrated a clear advantage of adding genomic analysis to risk assessment. There is little overlapping among signatures and merging their prognostic potential would be advantageous. Here, we describe a new strategy to merge published neuroblastoma related gene signatures into a single, highly accurate, Multi-Signature Ensemble (MuSE)-classifier of neuroblastoma (NB) patients outcome.

Biocompatibility and Antifibrotic Effect of UV-cross-linked Hyaluronate As a Release-system for Tranilast After Trabeculectomy in a Rabbit Model--a Pilot Study

To analyze the release kinetics and the clinical and histological effects of UV-cross-linked hyaluronic acid as a release-system for the transforming growth factor β-2 antagonist tranilast with anti-phlogistic properties on intraocular pressure after trabeculectomy in an aggressive scarring animal model.

Knowledge-based Potential for Positioning Membrane-associated Structures and Assessing Residue-specific Energetic Contributions

The complex hydrophobic and hydrophilic milieus of membrane-associated proteins pose experimental and theoretical challenges to their understanding. Here, we produce a nonredundant database to compute knowledge-based asymmetric cross-membrane potentials from the per-residue distributions of C(β), C(γ) and functional group atoms. We predict transmembrane and peripherally associated regions from genomic sequence and position peptides and protein structures relative to the bilayer (available at http://www.degradolab.org/ez). The pseudo-energy topological landscapes underscore positional stability and functional mechanisms demonstrated here for antimicrobial peptides, transmembrane proteins, and viral fusion proteins. Moreover, experimental effects of point mutations on the relative ratio changes of dual-topology proteins are quantitatively reproduced. The functional group potential and the membrane-exposed residues display the largest energetic changes enabling to detect native-like structures from decoys. Hence, focusing on the uniqueness of membrane-associated proteins and peptides, we quantitatively parameterize their cross-membrane propensity, thus facilitating structural refinement, characterization, prediction, and design.

Routinization and Medicalization of Palliative Care: Losses, Gains and Challenges

This article investigates some of the criticisms that have been directed at the hospice movement in the process of interaction with the traditional Western healthcare system, such as those relative to its routinization and medicalization. It also aims to review some of the consequences of this process of institutionalisation for the field of end-of-life care: surveillance and control over the process of dying, at the expense of decisions preferably based on the patient and that patient's ability to decide how to die, with the loss of wider objectives originally established by the movement, such as unconditional reception for the patient. Based on these criticisms, some considerations are made regarding the moral implications and risks related to this specific mode of action, the hospice way of care.

Pharmacological Activation of the P53 Pathway by Nutlin-3 Exerts Anti-tumoral Effects in Medulloblastomas

Medulloblastomas account for 20% of pediatric brain tumors. With an overall survival of 40%-70%, their treatment is still a challenge. The majority of medulloblastomas lack p53 mutations, but even in cancers retaining wild-type p53, the tumor surveillance function of p53 is inhibited by the oncoprotein MDM2. Deregulation of the MDM2/p53 balance leads to malignant transformation. Here, we analyzed MDM2 mRNA and protein expression in primary medulloblastomas and normal cerebellum and assessed the mutational status of p53 and MDM2 expression in 6 medulloblastoma cell lines. MDM2 expression was elevated in medulloblastomas, compared with cerebellum. Four of 6 medulloblastoma cell lines expressed wild-type p53 and high levels of MDM2. The tumor-promoting p53-MDM2 interaction can be inhibited by the small molecule, nutlin-3, restoring p53 function. Targeting the p53-MDM2 axis using nutlin-3 significantly reduced cell viability and induced either cell cycle arrest or apoptosis and expression of the p53 target gene p21 in these 4 cell lines. In contrast, DAOY and UW-228 cells harboring TP53 mutations were almost unaffected by nutlin-3 treatment. MDM2 knockdown in medulloblastoma cells by siRNA mimicked nutlin-3 treatment, whereas expression of dominant negative p53 abrogated nutlin-3 effects. Oral nutlin-3 treatment of mice with established medulloblastoma xenografts inhibited tumor growth and significantly increased survival. Thus, nutlin-3 reduced medulloblastoma cell viability in vitro and in vivo by re-activating p53 function. We suggest that inhibition of the MDM2-p53 interaction with nutlin-3 is a promising therapeutic option for medulloblastomas with functional p53 that should be further evaluated in clinical trials.

Role of Psychotherapy in the Management of Psychiatric Diseases

Psychotherapy plays an essential role in the treatment of mental disorders. The use and research of psychological treatment strategies increased drastically over the past decade. The general efficacy of psychotherapy for the treatment of psychiatric diseases is proved and documented in several meta-analyses. Psychotherapy re-searchers have found solutions for acceptable study designs which account for the special character of these interventions and studied the efficacy of psychotherapeutic treatment in more than 1000 intervention trials.Meanwhile evidence-based psychotherapy approaches tailored to a specific diagnosis are dominating the field and question the basis of psychotherapy schools.A new field of research in psychotherapy is the neurobiological basis of mental disorders and the demonstration of neurobiological changes with psycho-therapeutic treatment.

Role of Ultrasound Compared to Age-related Formulas for Uncuffed Endotracheal Intubation in a Pediatric Population

It is often difficult to determine the correct size of endotracheal tubes (ETT) needed for intubating pediatric patients. Therefore, we evaluated the role of ultrasound in pediatric patients to compare the correct size of an uncuffed (ETT) with the minimal transverse diameter of the subglottic airway (MTDSA) measured by ultrasound and with tube size predicted by different age-related formulas.

[Importance of PET/CT for Imaging of Colorectal Cancer]

CLINICAL/METHODICAL ISSUE: Fluorodeoxyglucose-positron emission tomography/computed tomography (FDG-PET/CT) has emerged as a very useful imaging modality in the management of colorectal carcinoma. Data from the literature regarding the role of PET/CT in the initial diagnosis, staging, radiotherapy planning, response monitoring and surveillance of colorectal carcinoma is presented. Future directions and economic aspects are discussed. STANDARD RADIOLOGICAL METHODS: Computed tomography (CT), magnetic resonance imaging (MRI) and FDG-PET for colorectal cancer and endorectal ultrasound for rectal cancer. METHODICAL INNOVATIONS: Combined FDG-PET/CT. PERFORMANCE: While other imaging modalities allow superior visualization of the extent and invasion depth of the primary tumor, PET/CT is most sensitive for the detection of distant metastases of colorectal cancer. ACHIEVEMENTS: We recommend a targeted use of PET/CT in cases of unclear M staging, prior to metastasectomy and in suspected cases of residual or recurrent colorectal carcinoma with equivocal conventional imaging. The role of PET/CT in radiotherapy planning and response monitoring needs to be determined. Currently there is no evidence to support the routine use of PET/CT for colorectal screening, staging or surveillance. PRACTICAL RECOMMENDATIONS: To optimally exploit the synergy between morphologic and functional information, FDG-PET should generally be performed as an integrated FDG-PET/CT with a contrast-enhanced CT component in colorectal carcinoma.

Haemophilia Care in Europe: the ESCHQoL Study

The aim of this study was to determine the clinical conditions of patients with haemophilia within Europe as recommended by the European Commission. In this multicentre, cross-sectional, ambispective study, conducted within 21 European countries patients' clinical data were collected, amongst others haemophilia type, severity, treatment pattern, use of factor products, bleeding, orthopaedic joint scores and infections. A total of 1400 patients, 84.3% with haemophilia A and 15.7% with haemophilia B were enrolled by 42 centres between 2004 and 2006. Thereof, 417 were children (30.0%) and 983 were adults (70.0%). About 70% of patients had severe factor deficiency (<1%). More than half of the adults were carriers of chronic infections (12.6% HIV, 55.8% HCV), compared to only 3.8% children (no HIV, 2.9% HCV). Patients were grouped according to per capita amount of clotting factor used in patients' region of residence in 2005: region 1: >5 IU; region 2: 2-5 IU; region 3: <2 IU. Paediatric and adult patients in region 3 had median numbers of three and eight joint bleeds, respectively, with worse joint scores compared to region 1 with zero and one bleed. Prophylactic therapy was used in only 31.3% children and 8.9% adults with severe haemophilia in region 3 compared to 93.7% and 54.1%, respectively, in region 1. Statistical analysis revealed that residence in areas with low factor consumption/availability is the most prominent risk factor for joint disease. Access of European patients with haemophilia to optimal care with safe factor VIII concentrates is limited and depends on the region of residence.

Field Evaluation of a Simple Fluorescence Method for Detection of Viable Mycobacterium Tuberculosis in Sputum Specimens During Treatment Follow-up

Simple tuberculosis (TB) treatment monitoring tools are needed. We assessed the performance of fluorescein-diacetate (FDA) smear microscopy for detection of viable Mycobacterium tuberculosis in sputum specimens (n = 288) of TB cases under treatment compared to culture (17.4% culture positivity). FDA sensitivity was moderate (83.7% [95% confidence interval {CI}, 70.3 to 92.6]), and specificity was low (66.1% [59.5 to 72.2]). The good negative predictive value (94.8% [90.1 to 97.8]) and negative likelihood ratio (0.2) suggest using this method to rule out treatment failure in settings without access to culture.

Intra- and Interprotein Phosphorylation Between Two-hybrid Histidine Kinases Controls Myxococcus Xanthus Developmental Progression

Histidine-aspartate phosphorelay signaling systems are used to couple stimuli to cellular responses. A hallmark feature is the highly modular signal transmission modules that can form both simple "two-component" systems and sophisticated multicomponent systems that integrate stimuli over time and space to generate coordinated and fine-tuned responses. The deltaproteobacterium Myxococcus xanthus contains a large repertoire of signaling proteins, many of which regulate its multicellular developmental program. Here, we assign an orphan hybrid histidine protein kinase, EspC, to the Esp signaling system that negatively regulates progression through the M. xanthus developmental program. The Esp signal system consists of the hybrid histidine protein kinase, EspA, two serine/threonine protein kinases, and a putative transport protein. We demonstrate that EspC is an essential component of this system because ΔespA, ΔespC, and ΔespA ΔespC double mutants share an identical developmental phenotype. Neither substitution of the phosphoaccepting histidine residue nor deletion of the entire catalytic ATPase domain in EspC produces an in vivo mutant developmental phenotype. In contrast, substitution of the receiver phosphoaccepting residue yields the null phenotype. Although the EspC histidine kinase can efficiently autophosphorylate in vitro, it does not act as a phosphodonor to its own receiver domain. Our in vitro and in vivo analyses suggest the phosphodonor is instead the EspA histidine kinase. We propose EspA and EspC participate in a novel hybrid histidine protein kinase signaling mechanism involving both inter- and intraprotein phosphotransfer. The output of this signaling system appears to be the combined phosphorylated state of the EspA and EspC receiver modules. This system regulates the proteolytic turnover of MrpC, an important regulator of the developmental program.

Older Drivers' Crashes in Queensland, Australia

The growing proportion of older adults in Australia is predicted to comprise 23% of the population by 2030. Accordingly, an increasing number of older drivers and fatal crashes of these drivers could also be expected. While the cognitive and physiological limitations of ageing and their road safety implications have been widely documented, research has generally considered older drivers as a homogeneous group. Knowledge of age-related crash trends within the older driver group itself is currently limited.

An Unusual Cause of Hypertriglyceridemic Acute Pancreatitis

Current Research with Cochlear Implants at Arizona State University

In this article we review, and discuss the clinical implications of, five projects currently underway in the Cochlear Implant Laboratory at Arizona State University. The projects are (1) norming the AzBio sentence test, (2) comparing the performance of bilateral and bimodal cochlear implant (CI) patients in realistic listening environments, (3) accounting for the benefit provided to bimodal patients by low-frequency acoustic stimulation, (4) assessing localization by bilateral hearing aid patients and the implications of that work for hearing preservation patients, and (5) studying heart rate variability as a possible measure for quantifying the stress of listening via an implant. The long-term goals of the laboratory are to improve the performance of patients fit with cochlear implants and to understand the mechanisms, physiological or electronic, that underlie changes in performance. We began our work with cochlear implant patients in the mid-1980s and received our first grant from the National Institutes of Health (NIH) for work with implanted patients in 1989. Since that date our work with cochlear implant patients has been funded continuously by the NIH. In this report we describe some of the research currently being conducted in our laboratory.

Malaria Parasite Type 4 Equilibrative Nucleoside Transporters (ENT4) Are Purine Transporters with Distinct Substrate Specificity

Malaria, caused by Plasmodia parasites, affects hundreds of millions of people. As purine auxotrophs, Plasmodia use transporters to import host purines for subsequent metabolism by the purine salvage pathway. Thus purine transporters are attractive drug targets. All sequenced Plasmodia genomes encode four ENTs (equilibrative nucleoside transporters). During the pathogenic intraerythrocytic stages, ENT1 is a major route of purine nucleoside/nucleobase transport. Another plasma membrane purine transporter exists because Plasmodium falciparum ENT1-knockout parasites survive at supraphysiological purine concentrations. The other three ENTs have not been characterized functionally. Codon-optimized Pf- (P. falciparum) and Pv- (Plasmodium vivax) ENT4 were expressed in Xenopus laevis oocytes and substrate transport was determined with radiolabelled substrates. ENT4 transported adenine and 2'-deoxyadenosine at the highest rate, with millimolar-range apparent affinity. ENT4-expressing oocytes did not accumulate hypoxanthine, a key purine salvage pathway substrate, or AMP. Micromolar concentrations of the plant hormone cytokinin compounds inhibited both PfENT4 and PvENT4. In contrast with PfENT1, ENT4 interacted with the immucillin compounds in the millimolar range and was inhibited by 10 μM dipyridamole. Thus ENT4 is a purine transporter with unique substrate and inhibitor specificity. Its role in parasite physiology remains uncertain, but is likely to be significant because of the strong conservation of ENT4 homologues in Plasmodia genomes.

Specific Glycan Elements Determine Differential Binding of Individual Egg Glycoproteins of the Human Parasite Schistosoma Mansoni by Host C-type Lectin Receptors

During infection with the blood fluke Schistosoma mansoni, glycan motifs present on glycoproteins of the parasite's eggs mediate immunomodulatory effects on the host. The recognition of these glycan motifs is primarily mediated by C-type lectin receptors on dendritic cells and other cells of the immune system. However, it is not yet known which individual glycoproteins interact with the different C-type lectin receptors, and which structural components are involved. Here we investigated the structural basis of the binding of two abundant egg antigens, kappa-5 and IPSE/α1, by the C-type lectin receptor dendritic cell-specific ICAM3-grabbing non-integrin, macrophage galactose-type lectin and mannose receptor. In the natural soluble form, the secretory egg glycoprotein IPSE/α1 interacts with dendritic cells mainly via mannose receptors. Surprisingly, in plate-based assays mannose receptors preferentially bound to mannose conjugates, while in cell-based assays, IPSE/α1 is bound via the fucosylated Galβ1-4(Fucα1-3)GlcNAc (LeX) motif on diantennary N-glycans. Kappa-5, in contrast, is bound by dendritic cells via all three C-type lectin receptors studied and for a minor part also via other, non-C-type lectin receptors. Kappa-5 interacts with macrophage galactose-type lectins via the GalNAcβ1-4GlcNAc antenna present on its triantennary N-glycans, as well as the GalNAcβ1-4(Fucα1-3)GlcNAc antennae present on a minor N-glycan subset. Dendritic cell-specific ICAM3-grabbing non-integrin binding of kappa-5 was mediated via the GalNAcβ1-4(Fucα1-3)GlcNAc antennae, whereas binding of mannose receptors may involve either GalNAcβ1-4(Fucα1-3)GlcNAc antennae or the fucosylated and xylosylated chitobiose core. This study provides a molecular and structural basis for future studies of the interaction between C-type lectin receptors and other soluble egg antigen glycoproteins and their effects on the host immune response.

Direct Observation of Prompt Pre-thermal Laser Ion Sheath Acceleration

High-intensity laser plasma-based ion accelerators provide unsurpassed field gradients in the megavolt-per-micrometer range. They represent promising candidates for next-generation applications such as ion beam cancer therapy in compact facilities. The weak scaling of maximum ion energies with the square-root of the laser intensity, established for large sub-picosecond class laser systems, motivates the search for more efficient acceleration processes. Here we demonstrate that for ultrashort (pulse duration ~30 fs) highly relativistic (intensity ~10(21) W cm(-2)) laser pulses, the intra-pulse phase of the proton acceleration process becomes relevant, yielding maximum energies of around 20 MeV. Prominent non-target-normal emission of energetic protons, reflecting an engineered asymmetry in the field distribution of promptly accelerated electrons, is used to identify this pre-thermal phase of the acceleration. The relevant timescale reveals the underlying physics leading to the near-linear intensity scaling observed for 100 TW class table-top laser systems.

Routinization and Medicalization of Palliative Care: Losses, Gains and Challenges-CORREGENDUM

Assessment of Therapeutic Options for Mild Obstructive Sleep Apnea Using Cardiopulmonary Coupling Measures

To examine the efficacy of various therapeutic modalities for mild obstructive sleep apnea using cardiopulmonary coupling variables of sleep quality.

Seizure Outcome, Functional Outcome, and Quality of Life After Hemispherectomy in Adults

Functional hemispherectomy is a well-established method in childhood epilepsy surgery with only a few reports on its application in adults.

The Influence of Temperature on the Photoluminescence Properties of Single InAs Quantum Dots Grown on Patterned GaAs

We report the temperature-dependent photoluminescence of single site-controlled and self-assembled InAs quantum dots. We have used nanoimprint lithography for patterning GaAs(100) templates and molecular beam epitaxy for quantum dot deposition. We show that the influence of the temperature on the photoluminescence properties is similar for quantum dots on etched nanopatterns and randomly positioned quantum dots on planar surfaces. The photoluminescence properties indicate that the prepatterning does not degrade the radiative recombination rate for the site-controlled quantum dots.

Regulatory B Cells from Hilar Lymph Nodes of Tolerant Mice in a Murine Model of Allergic Airway Disease Are CD5(+), Express TGF-β, and Co-localize with CD4(+)Foxp3(+) T Cells

In a biphasic, ovalbumin (OVA)-induced murine asthma model where allergic airway disease is followed by resolution and the development of local inhalational tolerance (LIT), transforming growth factor (TGF)-β-expressing CD5(+) B cells were selectively expanded locally in hilar lymph nodes (HLN) of LIT mice. LIT HLN CD5(+) B cells, but not LIT HLN CD5(-) B cells, induced expression of Foxp3 in CD4(+)CD25(-) T cells in vitro. These CD5(+) regulatory B cells (Breg) and CD4(+)Foxp3(+) T cells demonstrated similar increases in expression of chemokine receptors (CXCR4 and CXCR5) and co-localized in HLN B cell zones of LIT mice. The adoptive transfer of LIT HLN CD5(+) B cells, but not LIT HLN CD5(-) B cells, increased the number of CD4(+)Foxp3(+) T cells in the lung and inhibited airway eosinophilia in this OVA model. Thus, Breg in HLNs of LIT mice reside in a CD5(+) TGF-β-producing subpopulation and co-localize with CD4(+)Foxp3(+) T cells.

Chronic Autophagy is a Cellular Adaptation to Tumor Acidic PH Microenvironments

Tumor cell survival relies upon adaptation to the acidic conditions of the tumor microenvironment. To investigate potential acidosis survival mechanisms, we examined the effect of low pH (6.7) on human breast carcinoma cells. Acute low pH exposure reduced proliferation rate, induced a G1 cell cycle arrest, and increased cytoplasmic vacuolization. Gene expression analysis revealed elevated levels of ATG5 and BNIP3 in acid-conditioned cells, suggesting cells exposed to low pH may utilize autophagy as a survival mechanism. In support of this hypothesis, we found that acute low pH stimulated autophagy as defined by an increase in LC3-positive punctate vesicles, double-membrane vacuoles, and decreased phosphorylation of AKT and ribosomal protein S6. Notably, cells exposed to low pH for approximately 3 months restored their proliferative capacity while maintaining the cytoplasmic vacuolated phenotype. Although autophagy is typically transient, elevated autophagy markers were maintained chronically in low pH conditioned cells as visualized by increased protein expression of LC3-II and double-membrane vacuoles. Furthermore, these cells exhibited elevated sensitivity to PI3K-class III inhibition by 3-methyladenine. In mouse tumors, LC3 expression was reduced by systemic treatment with sodium bicarbonate, which raises intratumoral pH. Taken together, these results argue that acidic conditions in the tumor microenvironment promote autophagy, and that chronic autophagy occurs as a survival adaptation in this setting.

Seizure Outcome 1 year After Temporal Lobe Epilepsy: an Analysis of MR Volumetric and Clinical Parameters

The aim of this work was to determine predictors that may contribute to surgical success or failure. Relevant pre- and postoperative baseline data were analyzed, and temporal structures underwent a volumetric analysis.

Acyclic Immucillin Phosphonates: Second-generation Inhibitors of Plasmodium Falciparum Hypoxanthine-guanine-xanthine Phosphoribosyltransferase

Plasmodium falciparum, the primary cause of deaths from malaria, is a purine auxotroph and relies on hypoxanthine salvage from the host purine pool. Purine starvation as an antimalarial target has been validated by inhibition of purine nucleoside phosphorylase. Hypoxanthine depletion kills Plasmodium falciparum in cell culture and in Aotus monkey infections. Hypoxanthine-guanine-xanthine phosphoribosyltransferase (HGXPRT) from P. falciparum is required for hypoxanthine salvage by forming inosine 5'-monophosphate, a branchpoint for all purine nucleotide synthesis in the parasite. Here, we present a class of HGXPRT inhibitors, the acyclic immucillin phosphonates (AIPs), and cell permeable AIP prodrugs. The AIPs are simple, potent, selective, and biologically stable inhibitors. The AIP prodrugs block proliferation of cultured parasites by inhibiting the incorporation of hypoxanthine into the parasite nucleotide pool and validates HGXPRT as a target in malaria.

Pulmonary Mechanics Following Albuterol Therapy in Mechanically Ventilated Infants with Bronchiolitis

Bronchiolitis is a common cause of critical illness in infants. Inhaled β(2)-agonist bronchodilators are frequently used as part of treatment, despite unproven effectiveness. The purpose of this study was to describe the physiologic response to these medications in infants intubated and mechanically ventilated for bronchiolitis.

Creatine Increases Hippocampal Na(+),K(+)-ATPase Activity Via NMDA-calcineurin Pathway

Achievements made over the past few years have demonstrated the important role of the creatine and phosphocreatine system in the buffering and transport of high-energy phosphates into the brain; however, the non-energetic processes elicited by this guanidine compound in the hippocampus are still poorly understood. In the present study we disclosed that the incubation of rat hippocampal slices with creatine (10mM) for 30 min increased Na(+),K(+)-ATPase activity. In addition, intrahippocampal injection of creatine (5 nmol/site) also increased the above-mentioned activity. The incubation of hippocampal slices with N-methyl-d-aspartate (NMDA; MK-801, 10 μM) and NMDA Receptor 2B (NR2B; ifenprodil, 3 μM) antagonists but not with the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPA)/kainate antagonist (DNQX, 10 μM) and nitric oxide synthase inhibitor (NOS; l-NAME, 100 μM), blunted the effect of creatine on Na(+),K(+)-ATPase activity. Furthermore, the calcineurin inhibitor (cyclosporine A, 200 nM) as well as the Protein Kinase C (PMA, 100 nM) and Protein Kinase A (8-Br-cAMP, 30 μM) activators attenuated the creatine-induced increase of Na(+),K(+)-ATPase activity. In addition, the incubation of hippocampal slices with creatine (10mM) for 30 min increased calcineurin activity. The results presented here suggest that creatine increases Na(+),K(+)-ATPase activity via NMDA-calcineurin pathway, proposing an putative underlying non-energetic role of this guanidine compound. However, more studies are needed to assess the contribution of this putative alternative role in neurological diseases that present decreased Na(+),K(+)-ATPase activity.

Targeted Expression of Mutated ALK Induces Neuroblastoma in Transgenic Mice

Activating anaplastic lymphoma kinase (ALK) mutations were recently detected in most familial and 10% of sporadic neuroblastomas. However, the role of mutated ALK in tumorigenesis remains elusive. We demonstrate that targeted expression of the most frequent and aggressive variant, ALK(F1174L), is tumorigenic in mice. Tumors resembled human neuroblastomas in morphology, metastasis pattern, gene expression, and the presence of neurosecretory vesicles as well as synaptic structures. This ALK-driven neuroblastoma mouse model precisely recapitulated the genetic spectrum of the disease. Chromosomal aberrations were syntenic to those in human neuroblastoma, including 17q gain and MYCN oncogene amplification. Targeted ALK(F1174L) and MYCN coexpression revealed a strong synergism in inducing neuroblastoma with minimal chromosomal aberrations, suggesting that fewer secondary hits are required for tumor induction if both oncoproteins are targeted. Treatment of ALK(F1174L) transgenic mice with the ALK inhibitor TAE-684 induced complete tumor regression, indicating that tumor cells were addicted to ALK(F1174L) activity. We conclude that an activating mutation within the ALK kinase domain is sufficient to induce neuroblastoma development, and ALK inhibitors show promise for treating human neuroblastomas harboring ALK mutations.

Microarray Analyses of Inflammation Response of Human Dermal Fibroblasts to Different Strains of Borrelia Burgdorferi Sensu Stricto

In Lyme borreliosis, the skin is the key site of bacterial inoculation by the infected tick, and of cutaneous manifestations, erythema migrans and acrodermatitis chronica atrophicans. We explored the role of fibroblasts, the resident cells of the dermis, in the development of the disease. Using microarray experiments, we compared the inflammation of fibroblasts induced by three strains of Borrelia burgdorferi sensu stricto isolated from different environments and stages of Lyme disease: N40 (tick), Pbre (erythema migrans) and 1408 (acrodermatitis chronica atrophicans). The three strains exhibited a similar profile of inflammation with strong induction of chemokines (CXCL1 and IL-8) and IL-6 cytokine mainly involved in the chemoattraction of immune cells. Molecules such as TNF-alpha and NF-κB factors, metalloproteinases (MMP-1, -3 and -12) and superoxide dismutase (SOD2), also described in inflammatory and cellular events, were up-regulated. In addition, we showed that tick salivary gland extracts induce a cytotoxic effect on fibroblasts and that OspC, essential in the transmission of Borrelia to the vertebrate host, was not responsible for the secretion of inflammatory molecules by fibroblasts. Tick saliva components could facilitate the early transmission of the disease to the site of injury creating a feeding pit. Later in the development of the disease, Borrelia would intensively multiply in the skin and further disseminate to distant organs.

Frozen Section Diagnosis in Donor Liver Biopsies: Observer Variation of Semiquantitative and Quantitative Steatosis Assessment

Donor livers are not generally accepted for liver transplantation if intraoperative frozen section histology on wedge biopsies provides evidence for more severe steatosis. In this reliability study, assessment of steatosis in donor liver biopsies by different approaches (frozen sections vs. paraffin sections; macrovesicular steatosis vs. microvesicular steatosis), different observers, and different evaluation methods (conventional microscopy vs. point grid analysis on digital microphotographs) was compared. One hundred twenty consecutive donor liver biopsies were investigated. Intraoperative diagnosis was made on hematoxylin and eosin (H&E)-stained frozen sections. The residual portion of each biopsy was analyzed later on H&E-, diastase-resistant PAS-, and Elastica van Gieson-stained paraffin sections. Microvesicular steatosis and macrovesicular steatosis were classified semiquantitatively into 5 % steps. Additionally, point grid counting was applied on ten digital microphotographs per slide. The values for steatosis revealed a wide range of data between 0 and 70 or 85 % (mean values, 12.0-18.3 %), considering all types of specimens. The results of the two observers were highly correlated for macrovesicular steatosis (r ≥ 0.925) and for microvesicular steatosis (r ≥ 0.880). The values for macrovesicular and microvesicular steatosis, however, showed poor correlation (r ≤ 0.581). The rate of agreement between the two observers ranged between 84.2 and 95.8 % (κ, 0.763-0.937), depending on the threshold setting. For point grid analysis, significantly lower mean values and ranges for both types of steatosis compared to conventional histopathology were found (p < 0.001). Comparing the results of point grid analysis with those of conventional histopathology, a relatively loose correlation was found (r, 0.581-0.779). Intraoperative histology remains a reliable and highly relevant method for the assessment of steatosis in liver donor grafts. It represents one important component in the decision-finding whether or not a donor liver should be accepted and should possibly be combined with results of preoperative computed tomography imaging. Considering our data, macrovesicular and microvesicular steatosis should be analyzed separately due to the limited correlation between them.

Neuropsychological Outcome After Selective Amygdalohippocampectomy: Subtemporal Versus Transsylvian Approach

In the context of discussions on the optimal cognitive outcome of temporal lobe epilepsy surgery, and stimulated by recent reports on the beneficial effects of a selective subtemporal approach to memory function, this study evaluated the cognitive consequences of subtemporal versus transsylvian selective amygdalohippocampectomy (SAH) in patients with mesial temporal lobe epilepsy, taking verbal/figural memory and language functions into account.

β2-adrenergic Receptor Haplotype Linked to Intubation and Mechanical Ventilation in Children with Asthma

Children with asthma and respiratory failure comprise a small but significant subset of children with acute asthma. In addition to clinical and historical factors that have been associated with respiratory failure, there may also be genetic factors that predispose some asthmatic children to intubation and mechanical ventilation. However, this has not previously been assessed in this population. We hypothesized that genetic polymorphisms of the β(2)-adrenergic receptor (ADRβ(2)) are associated with intubation and mechanical ventilation in children with asthma.

Modulation of Neuroblastoma Disease Pathogenesis by an Extensive Network of Epigenetically Regulated MicroRNAs

MicroRNAs (miRNAs) contribute to the pathogenesis of many forms of cancer, including the pediatric cancer neuroblastoma, but the underlying mechanisms leading to altered miRNA expression are often unknown. Here, a novel integrated approach for analyzing DNA methylation coupled with miRNA and mRNA expression data sets identified 67 epigenetically regulated miRNA in neuroblastoma. A large proportion (42%) of these miRNAs was associated with poor patient survival when underexpressed in tumors. Moreover, we demonstrate that this panel of epigenetically silenced miRNAs targets a large set of genes that are overexpressed in tumors from patients with poor survival in a highly redundant manner. The genes targeted by the epigenetically regulated miRNAs are enriched for a number of biological processes, including regulation of cell differentiation. Functional studies involving ectopic overexpression of several of the epigenetically silenced miRNAs had a negative impact on neuroblastoma cell viability, providing further support to the concept that inactivation of these miRNAs is important for neuroblastoma disease pathogenesis. One locus, miR-340, induced either differentiation or apoptosis in a cell context dependent manner, indicating a tumor suppressive function for this miRNA. Intriguingly, it was determined that miR-340 is upregulated by demethylation of an upstream genomic region that occurs during the process of neuroblastoma cell differentiation induced by all-trans retinoic acid (ATRA). Further biological studies of miR-340 revealed that it directly represses the SOX2 transcription factor by targeting of its 3'-untranslated region, explaining the mechanism by which SOX2 is downregulated by ATRA. Although SOX2 contributes to the maintenance of stem cells in an undifferentiated state, we demonstrate that miR-340-mediated downregulation of SOX2 is not required for ATRA induced differentiation to occur. In summary, our results exemplify the dynamic nature of the miRNA epigenome and identify a remarkable network of miRNA/mRNA interactions that significantly contribute to neuroblastoma disease pathogenesis.Oncogene advance online publication, 16 July 2012; doi:10.1038/onc.2012.311.

Impacts of Climate-induced Changes on the Distribution of Pesticides Residues in Water and Sediment of Lake Naivasha, Kenya

This study reports evidence of increased chlorpyrifos contamination in sediment and water in Lake Naivasha following its intensive application in the horticultural farms in the catchment area. Analytical results show that levels of chlorpyrifos residues were influenced by climate-induced rainfall pattern with higher levels reported during period of heavy precipitation with significant decrease during low rainfall. On average, the levels ranged between 14.8 and 32.8 ng g(-1) in sediment during rainy season compared to a range of 8.5-16.6 ng g(-1) in the dry season. Additionally, the mean concentration of chlorpyrifos in water ranged between 8.61 and 22.4 μg L(-1) during rainy season and below detection limit (bdl) -13.6 μg L(-1) in dry season as quantified by enzyme-linked immunosorbent assay. Meanwhile, independent t test analysis indicated that there was significant difference in concentration at p ≤ 0.05 between the seasons with respect to sediment and water samples. This demonstrated that climate-induced variations had considerable influence on contamination. While diazinon and carbofuran were equally applied intensively, their levels were below the detection limit in the all the samples analyzed. ELISA results were validated by the capillary-HPLC photodiode-array detector instrument analysis, and statistical comparison showed no significant difference between them. It was evident that chlorpyrifos residues determination in water and sediment by ELISA can be a useful strategy in environmental management and monitoring program, and a complimentary analytical tool to high performance liquid chromatography. Levels of chlorpyrifos detected in sediment and water were found to exceed recommended criteria for protection of aquatic life and preservation of water quality and may be hazardous if not regularly monitored.

[Autoimmune Liver Diseases]

Autoimmune liver diseases are divided into autoimmune hepatitis (AIH), primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC). They play an important role in the differential diagnosis of acute and chronic liver diseases. Diagnostic criteria consist of biochemical parameters (liver function tests, immunoglobulins, autoantibodies), imaging (ultrasound, MRCP, ERCP) and histopathologic characteristics. The treatment of choice in AIH is a combined immunosuppressive therapy of prednisolone and azathioprine. Ursodeoxycholic acid is the treatment of choice for PBC and improves liver biochemistry and prolongs transplant-free survival by slowing histological progression in the majority of patients. To date, there is no effective medical treatment option for patients with PSC.

Task Rules, Working Memory, and Fluid Intelligence

Many varieties of working memory have been linked to fluid intelligence. In Duncan et al. (Journal of Experimental Psychology:General 137:131-148, 2008), we described limited working memory for new task rules: When rules are complex, some may fail in their control of behavior, though they are often still available for explicit recall. Unlike other kinds of working memory, load is determined in this case not by real-time performance demands, but by the total complexity of the task instructions. Here, we show that the correlation with fluid intelligence is stronger for this aspect of working memory than for several other, more traditional varieties-including simple and complex spans and a test of visual short-term memory. Any task, we propose, requires construction of a mental control program that aids in segregating and assembling multiple task parts and their controlling rules. Fluid intelligence is linked closely to the efficiency of constructing such programs, especially when behavior is complex and novel.

Identification of the Spinal Pathways Involved in the Recovery of Baroreflex Control After Spinal Lesion in the Rat Using Pseudorabies Virus

Neurons in the rostroventrolateral medulla (RVLM) mediate baroreflex regulation (BR) of spinal sympathetic preganglionic neurons. Previously, our laboratory has shown that recovery of BR occurs in the rat after spinal hemisection. (Zahner MR, Kulikowicz E, and Schramm LP. Am J Physiol Regul Integr Comp Physiol 301: R1584-R1590, 2011). The goal of these experiments was to determine whether the observed recovery of BR is mediated by the reorganization of ipsilateral pathways or by compensation by spared contralateral pathways. To determine this, we infected the left kidney in rats with the retrograde transynaptic tracer, pseudorabies virus (PRV), either 1 or 8 wk after left spinal hemisection at either T(3) or T(8), or after a sham lesion. In sham-lesioned rats, PRV infection of RVLM neurons was bilateral. In all rats with a left hemisection, regardless of the location of the lesion (T(3) or T(8)) or postlesion recovery time (1 or 8 wk), PRV infection of left RVLM neurons was significantly reduced compared with sham-lesioned rats (P < 0.05). In a separate group of rats, we performed BR tests by measuring responses of left renal sympathetic nerve activity to pharmacologically induced decreases and increases in arterial pressure. In rats with T(8) left hemisection and 8-wk recovery, BR was robust, and acute right upper thoracic hemisection abolished all BR of left renal sympathetic nerve activity. Collectively, these data suggest that the recovery of BR is not mediated by reorganization of ipsilateral bulbospinal connections, but instead by improved efficacy of existing contralateral pathways.

High-throughput Quantitative Real-time Polymerase Chain Reaction Array for Absolute and Relative Quantification of Rhesus Macaque Types I, II, and III Interferon and Their Subtypes

Rhesus macaques provide a valuable research and preclinical model for cancer and infectious diseases, as nonhuman primates share immune pathways with humans. Interferons (IFNs) are key cytokines in both innate and adaptive immunity, so a detailed analysis of gene expression in peripheral blood and tissues may shed insight into immune responses. Macaques have 18 IFN genes, of which 14 encode for 13 distinct IFN-α subtypes, and one for IFN-β. Here, we developed a high-throughput array to evaluate each of the IFN-α subtypes, as well as IFN-β, IFN-γ and 2 subtypes of IFN-λ. With this array, expression of each IFN species may be quantified as relative to a reference (housekeeping) gene (ΔCq) or fitted to its own 4-point standard curve for absolute quantification (copy number per mass unit RNA). After validating the assay with IFN complementary DNA, we determined the IFN expression profile of peripheral blood mononuclear cells from 3 rhesus macaques in response to TLR agonists, and demonstrated that the profiles are consistent among animals. Furthermore, because the IFN expression profiles differ depending on the TLR stimuli, they suggest different biological functions for many of the IFN species measured, including individual subtypes of IFN-α.

Genome-wide Association Analysis in Sclerosing Cholangitis and Ulcerative Colitis Identifies Risk Loci at GPR35 and TCF4

Approximately 60-80% of patients with primary sclerosing cholangitis (PSC) have concurrent ulcerative colitis (UC). Previous genome-wide association studies (GWAS) in PSC have detected a number of susceptibility loci that also show associations in UC and other immune-mediated diseases. We aimed to systematically compare genetic associations in PSC with genotype data in UC patients with the purpose of detecting new susceptibility loci for PSC. We performed combined analyses of GWAS for PSC and UC comprising 392 PSC cases, 987 UC cases and 2,977 controls and followed up top association signals in additional 1,012 PSC cases, 4,444 UC cases and 11,659 controls. We discovered novel genome-wide significant associations with PSC at 2q37 [rs3749171 at GPR35; P=3.0×10(-9) in the overall study population, combined odds ratio (OR; 95% confidence interval (CI)) of 1.39 (1.24-1.55)], and at 18q21 [rs1452787 at TCF4; P=2.61×10(-8) , OR (95% CI) = 0.75 (0.68-0.83)]. In addition, several suggestive PSC associations were detected. The GPR35 rs3749171 is a missense single nucleotide polymorphism resulting in a shift from threonine to methionine. Structural modeling showed that rs3749171 is located in the third transmembrane helix of GPR35 and could possibly alter efficiency of signaling through the GPR35 receptor. Conclusion: By refining the analysis of a PSC GWAS by parallel assessments in a UC GWAS we were able to detect two novel risk loci at genome-wide significance levels. GPR35 shows associations in both UC and PSC, while TCF4 represents a PSC risk locus not associated with UC. Both loci may represent previously unexplored aspects of PSC pathogenesis. (HEPATOLOGY 2012.).

Progenitor Cell Homing in the Postischemic Myocardium: Just an Unmotivated Pitstop in the Microcirculation?

We developed a model for direct assessment of BMC sequestration in the postischemic murine myocardium after direct antegrade intracoronary injection.

Shell Biofilm-associated Nitrous Oxide Production in Marine Molluscs: Processes, Precursors and Relative Importance

Emission of the greenhouse gas nitrous oxide (N(2) O) from freshwater and terrestrial invertebrates has exclusively been ascribed to N(2) O production by ingested denitrifying bacteria in the anoxic gut of the animals. Our study of marine molluscs now shows that also microbial biofilms on shell surfaces are important sites of N(2) O production. The shell biofilms of Mytilus edulis, Littorina littorea and Hinia reticulata contributed 18-94% to the total animal-associated N(2) O emission. Nitrification and denitrification were equally important sources of N(2) O in shell biofilms as revealed by (15) N-stable isotope experiments with dissected shells. Microsensor measurements confirmed that both nitrification and denitrification can occur in shell biofilms due to a heterogeneous oxygen distribution. Accordingly, ammonium, nitrite and nitrate were important drivers of N(2) O production in the shell biofilm of the three mollusc species. Ammonium excretion by the animals was found to be sufficient to sustain N(2) O production in the shell biofilm. Apparently, the animals provide a nutrient-enriched microenvironment that stimulates growth and N(2) O production of the shell biofilm. This animal-induced stimulation was demonstrated in a long-term microcosm experiment with the snail H. reticulata, where shell biofilms exhibited the highest N(2) O emission rates when the animal was still living inside the shell.

Quantitative Measurement of Sleep Quality Using Cardiopulmonary Coupling Analysis: a Retrospective Comparison of Individuals with and Without Primary Insomnia

OBJECTIVE: The purpose of this study was to determine the utility of a new operator-independent, automated measure of sleep physiology based on cardiopulmonary coupling (CPC) analysis in subjects with primary insomnia vs. good sleepers. PATIENTS/METHODS: The polysomnograms of 50 subjects with primary insomnia and 36 good sleepers were summarized and analyzed from a consecutive two-night protocol. The electrocardiograms (ECG) from adaptation and baseline night polysomnograms were analyzed using CPC analysis. This Fourier-based technique uses heart rate variability and ECG R wave amplitude fluctuations associated with respiration to generate frequency maps of coupled autonomic-respiratory oscillations. The resulting sleep spectrogram is able to categorize sleep as "stable" (high-frequency coupling [HFC], 0.1-0.4 Hz) and "unstable" (low-frequency coupling [LFC], 0.1-0.01 Hz), independent of standard sleep stages. Wake and rapid eye movement sleep exhibit very low-frequency coupling (VLFC, 0.0039-0.01 Hz). Elevated LFC (e-LFC) is a subset of LFC that is associated with fragmented sleep of various etiologies. RESULTS: CPC variables showed a significant multivariate analysis of variance group, night, and group × night main effect, except for HFC by night. Relative to good sleepers, primary insomnia patients on adaptation night had lower HFC, a putative biomarker of stable sleep, and HFC/LFC ratio, an indicator of sleep quality. The primary insomnia group also had higher LFC, an index of unstable sleep, and an increase in VLFC and e-LFC compared to good sleepers on adaptation night. On baseline night, the primary insomnia group had increased LFC, VLFC, and e-LFC and a lower HFC/LFC ratio. Except for HFC, good sleepers had larger CPC variable differences between adaptation and baseline nights compared to the primary insomnia group. CONCLUSION: Primary insomnia subjects have a marked worsening of sleep quality on the adaptation night, which is well captured by both conventional and ECG-derived sleep spectrogram techniques. The larger improvement of sleep quality was found among good sleepers and captured only by CPC analysis. The operator-independent, automated measure of sleep physiology demonstrated functionality to differentiate and objectively quantify sleep quality.

ImzML--a Common Data Format for the Flexible Exchange and Processing of Mass Spectrometry Imaging Data

The application of mass spectrometry imaging (MS imaging) is rapidly growing with a constantly increasing number of different instrumental systems and software tools. The data format imzML was developed to allow the flexible and efficient exchange of MS imaging data between different instruments and data analysis software. imzML data is divided in two files which are linked by a universally unique identifier (UUID). Experimental details are stored in an XML file which is based on the HUPO-PSI format mzML. Information is provided in the form of a 'controlled vocabulary' (CV) in order to unequivocally describe the parameters and to avoid redundancy in nomenclature. Mass spectral data are stored in a binary file in order to allow efficient storage. imzML is supported by a growing number of software tools. Users will be no longer limited to proprietary software, but are able to use the processing software best suited for a specific question or application. MS imaging data from different instruments can be converted to imzML and displayed with identical parameters in one software package for easier comparison. All technical details necessary to implement imzML and additional background information is available at www.imzml.org.

Reply To: "Regulatory T Cell Defects in Autoimmune Hepatitis"

An Extreme Case of Plant-Insect Codiversification: Figs and Fig-Pollinating Wasps

It is thought that speciation in phytophagous insects is often due to colonization of novel host plants, because radiations of plant and insect lineages are typically asynchronous. Recent phylogenetic comparisons have supported this model of diversification for both insect herbivores and specialized pollinators. An exceptional case where contemporaneous plant-insect diversification might be expected is the obligate mutualism between fig trees (Ficus species, Moraceae) and their pollinating wasps (Agaonidae, Hymenoptera). The ubiquity and ecological significance of this mutualism in tropical and subtropical ecosystems has long intrigued biologists, but the systematic challenge posed by >750 interacting species pairs has hindered progress toward understanding its evolutionary history. In particular, taxon sampling and analytical tools have been insufficient for large-scale cophylogenetic analyses. Here, we sampled nearly 200 interacting pairs of fig and wasp species from across the globe. Two supermatrices were assembled: on an average, wasps had sequences from 77% of 6 genes (5.6 kb), figs had sequences from 60% of 5 genes (5.5 kb), and overall 850 new DNA sequences were generated for this study. We also developed a new analytical tool, Jane 2, for event-based phylogenetic reconciliation analysis of very large data sets. Separate Bayesian phylogenetic analyses for figs and fig wasps under relaxed molecular clock assumptions indicate Cretaceous diversification of crown groups and contemporaneous divergence for nearly half of all fig and pollinator lineages. Event-based cophylogenetic analyses further support the codiversification hypothesis. Biogeographic analyses indicate that the present-day distribution of fig and pollinator lineages is consistent with a Eurasian origin and subsequent dispersal, rather than with Gondwanan vicariance. Overall, our findings indicate that the fig-pollinator mutualism represents an extreme case among plant-insect interactions of coordinated dispersal and long-term codiversification. [Biogeography; coevolution; cospeciation; host switching; long-branch attraction; phylogeny.].

Transition State Analogue Inhibitors of Human Methylthioadenosine Phosphorylase and Bacterial Methylthioadenosine/S-adenosylhomocysteine Nucleosidase Incorporating Acyclic Ribooxacarbenium Ion Mimics

Several acyclic hydroxy-methylthio-amines with 3-5 carbon atoms were prepared and coupled via a methylene link to 9-deazaadenine. The products were tested for inhibition against human MTAP and Escherichia coli and Neisseria meningitidis MTANs and gave K(i) values as low as 0.23 nM. These results were compared to those obtained with 1st and 2nd generation inhibitors (1S)-1-(9-deazaadenin-9-yl)-1,4-dideoxy-1,4-imino-5-methylthio-D-ribitol (MT-Immucillin-A, 3) and (3R,4S)-1-[9-deazaadenin-9-yl)methyl]3-hydroxy-4-methylthiomethylpyrrolidine (MT-DADMe-Immucillin-A, 4). The best inhibitors were found to exhibit binding affinities of approximately 2- to 4-fold those of 3 but were significantly weaker than 4. Cleavage of the 2,3 carbon-carbon bond in MT-Immucillin-A (3) gave an acyclic product (79) with a 21,500 fold loss of activity against E. coli MTAN. In another case, N-methylation of a side chain secondary amine resulted in a 250-fold loss of activity against the same enzyme [(±)-65 vs (±)-68]. The inhibition results were also contrasted with those acyclic derivatives previously prepared as inhibitors for a related enzyme, purine nucleoside phosphorylase (PNP), where some inhibitors in the latter case were found to be more potent than their cyclic counterparts.

Signature of a Type-A Glass Transition and Intrinsic Confinement Effects in a Binary Glass-forming System

We study dynamically highly asymmetric binary mixtures comprised of small methyl tetrahydrofuran (MTHF) molecules and polystyrene. Combined use of dielectric spectroscopy, 2H nuclear magnetic resonance, incoherent quasielastic neutron scattering, and depolarized dynamic light scattering allows us to selectively probe the dynamics of the components in a broad dynamic range. It turns out that the mixtures exhibit two glass transitions in a wide concentration range although being fully miscible on a macroscopic scale. In between both glass transition temperatures, the dynamics of the small molecules show strong confinement effects, e.g., a crossover from Vogel-Fulcher to Arrhenius behavior of the time constants. Moreover, the dynamical behavior of small molecules close to the slow matrix is consistent with mode coupling theory predictions for a type-A glass transition, which was expected from recent theoretical and simulation studies in comparable systems.

Synthetic Lethality Between Rb, P53 and Dicer or MiR-17-92 in Retinal Progenitors Suppresses Retinoblastoma Formation

Synthetic lethality is a promising strategy for specific targeting of cancer cells that carry mutations that are absent in normal cells. This approach may help overcome the challenge associated with targeting dysfunctional tumour suppressors, such as p53 and Rb (refs 1, 2). Here we show that Dicer1 targeting prevents retinoblastoma formation in mice by synthetic lethality with combined inactivation of p53 and Rb. Although Dicer1 functions as a haploinsufficient tumour suppressor, its complete loss of function is selected against during tumorigenesis(3-5). We show that Dicer1 deficiency is tolerated in Rb-deficient retinal progenitor cells harbouring an intact p53 pathway, but not in the absence of p53. This synthetic lethality is mediated by the oncogenic miR-17-92 cluster because its deletion phenocopies Dicer1 loss in this context. miR-17-92 inactivation suppresses retinoblastoma formation in mice and co-silencing of miR-17/20a and p53 cooperatively decreases the viability of human retinoblastoma cells. These data provide an explanation for the selective pressure against loss of Dicer1 during tumorigenesis and a proof-of-concept that targeting miRNAs may potentially represent a general approach for synthetic lethal targeting of cancer cells that harbour specific cancer-inducing genotypes.

[Value of Magnetic Resonance Imaging in Diffuse Liver Diseases]

CLINICAL PROBLEM: Diffuse liver diseases show an increasing prevalence. The diagnostic gold standard of liver biopsy has several disadvantages. There is a clinical demand for non-invasive imaging-based techniques to qualitatively and quantitatively evaluate the entire liver. STANDARD RADIOLOGICAL METHODS: Ultrasound, computed tomography (CT) and magnetic resonance imaging (MRI) are routinely used. METHODICAL INNOVATIONS: Steatosis: chemical shift and frequency selective imaging, MR spectroscopy (MRS). Hemochromatosis: MR-based iron quantification. Fibrosis: MR elastography, diffusion, intravoxel incoherent motion (IVIM) and MR perfusion. PERFORMANCE/ACHIEVEMENTS/PRACTICAL RECOMMENDATIONS: T1-weighted in and opposed phase imaging is the clinically most frequently used MR technique to noninvasively detect and quantify steatosis. New methods for quantification that are not influenced by confounders like iron overload are under investigation. The most sensitive method to measure the fat content of the liver is MRS. As data acquisition and analysis remain complex and there is no whole organ coverage, MRS of the liver is not a routine method. With an optimized protocol incorporating T2* sequences, MRI is the modality of choice to quantify iron overload in hemochromatosis. Standard MR sequences cannot depict early stages of liver fibrosis. Advanced MR techniques (e.g. elastography, diffusion, IVIM and perfusion) for noninvasive assessment of liver fibrosis appear promising but their role has to be further investigated.

Constrained Bonding Environment in the Michaelis Complex of Trypanosoma Cruzi Uridine Phosphorylase

The transition state for the Trypanosoma cruzi uridine phosphorylase (TcUP) reaction has an expanded S(N)2 character. We used binding isotope effects (BIE's) to probe uridine distortion in the complex with TcUP and sulfate to mimic the Michaelis complex. Inverse 1'-(3)H and 5'-(3)H BIE's indicate a constrained bonding environment of these groups in the complex. Quantum chemical modeling identified a uridine conformer whose calculated BIE's match the experimental values. This conformer differs in sugar pucker and uracil orientation from the unbound conformer and the transition-state structure. These results support ground-state stabilization in the Michaelis complex.

A Picomolar Transition State Analogue Inhibitor of MTAN As a Specific Antibiotic for Helicobacter Pylori

Campylobacter and Helicobacter species express a 6-amino-6-deoxyfutalosine N-ribosylhydrolase (HpMTAN) proposed to function in menaquinone synthesis. BuT-DADMe-ImmA is a 36 pM transition state analogue of HpMTAN, and the crystal structure of the enzyme-inhibitor complex reveals the mechanism of inhibition. BuT-DADMe-ImmA has a MIC(90) value of <8 ng/mL for Helicobacter pylori growth but does not cause growth arrest in other common clinical pathogens, thus demonstrating potential as an H. pylori-specific antibiotic.

Dental Infection Simulating Skin Lesion

Orocutaneous fistulas or cutaneous sinus, a tract of dental origin, is an uncommon but well-documented condition that usually requires emergency treatment. Such condition may be misdiagnosed by physicians and dentists and may sometimes be confused with bone and skin tumor, osteomyelitis, congenital fistula, salivary gland fistula, pyogenic granuloma, infected cyst, deep mycotic infection, and other pathologies. A case of facial sinus tract that was initially misdiagnosed by a physician as a nonodontogenic lesion is presented. Nonsurgical endodontic therapy was the treatment of choice for this case. Facial cutaneous sinus tracts must be considered of dental origin. Early diagnosis and prompt treatment minimize patient discomfort and esthetic problems, reducing the possibility of further complications such as sepsis and osteomyelitis.

Bromelain Limits Airway Inflammation in an Ovalbumin-induced Murine Model of Established Asthma

Allergic asthma continues to increase despite new pharmacological advances for both acute treatment and chronic-disease management. Asthma is a multifactorial disease process with genetic, allergic, infectious, environmental, and dietary origins. Researchers are investigating the benefits of lifestyle changes and alternative asthma treatments, including the ability of bromelain to inhibit inflammation. Bromelain is a commonly used, proteolytically active pineapple extract.

A Truncated Lamin A in the Lmna -/- Mouse Line: Implications for the Understanding of Laminopathies

During recent years a number of severe clinical syndromes, collectively termed laminopathies, turned out to be caused by various, distinct mutations in the human LMNA gene. Arising from this, remarkable progress has been made to unravel the molecular pathophysiology underlying these disorders. A great benefit in this context was the generation of an A-type lamin deficient mouse line (Lmna (-/-) ) by Sullivan and others, ( 1) which has become one of the most frequently used models in the field and provided profound insights to many different aspects of A-type lamin function. Here, we report the unexpected finding that these mice express a truncated Lmna gene product on both transcriptional and protein level. Combining different approaches including mass spectrometry, we precisely define this product as a C-terminally truncated lamin A mutant that lacks domains important for protein interactions and post-translational processing. Based on our findings we discuss implications for the interpretation of previous studies using Lmna (-/-) mice and the concept of human laminopathies.

Acute Liver Failure Following Minocycline Treatment - a Case Report and Review of the Literature

We present the case of a 23-year-old female patient with acute liver failure following intake of minocycline. This patient had severe hypereosinophilia and massively increased IgE levels. Experimental studies in this case revealed elevated IFN-γ-, as well as TNF-α-producing CD4+ and CD8+ T-cells after in vitro stimulation with minocycline, indicating a type I/IgE-mediated as well as type II/cytotoxic reaction in the pathogenesis of minocycline-induced liver failure. Although mild forms of liver involvement are well known side effects of minocycline, only 8 cases with acute liver failure have been reported, and we present a review of all cases.

Pharmacological Characterization of Designer Cathinones in Vitro

Background and purpose: Designer β-keto amphetamines (e.g., cathinones, "bath salts," and "research chemicals") have become popular recreational drugs, but their pharmacology is poorly characterized. Experimental approach: We determined the potencies of cathinones to inhibit dopamine (DA), noradrenaline (NA), and serotonin (5-hydroxytryptamine [5-HT]) transport into transporter-transfected human embryonic kidney 293 cells, DA and 5-HT efflux from monoamine-preloaded cells, and monoamine receptor binding affinity. Key results: Mephedrone, methylone, ethylone, butylone, and naphyrone act as nonselective monoamine uptake inhibitors, similar to cocaine. Mephedrone, methylone, ethylone, and butylone also release 5-HT, similar to 3,4-methylenedioxymethamphetamine (MDMA, ecstasy) and other entactogens. Cathinone, methcathinone, and flephedrone act as preferential DA and NA uptake inhibitors and DA releasers, similar to amphetamine and methamphetamine. Pyrovalerone and 3,4-methylenedioxypyrovalerone (MDPV) are highly potent and selective DA and NA transporter inhibitors but unlike amphetamines do not release monoamines. The non-β-keto amphetamines are trace amine-associated receptor 1 ligands, whereas cathinones are not. All cathinones showed high blood-brain barrier permeability in an in vitro model. Mephedrone and MDPV exhibited particularly high permeability. Conclusions and implications: Cathinones have considerable pharmacological differences that form the basis for their suggested classification into three groups. The predominant action of all cathinones on the DA transporter is likely associated with a considerable risk of addiction. © 2012 The Authors. British Journal of Pharmacology © 2012 The British Pharmacological Society.

Metronidazole Immediate Release Formulations: a Fasting Randomized Open-label Crossover Bioequivalence Study in Healthy Volunteers

Metronidazole is a BCS (Biopharmaceutics Classification System) class 1 drug, traditionally considered the choice drug in the infections treatment caused by protozoa and anaerobic microorganisms. This study aimed to evaluate bioequivalence between 2 different marketed 250 mg metronidazole immediate release tablets. A randomized, open-label, 2×2 crossover study was performed in healthy Brazilian volunteers under fasting conditions with a 7-day washout period. The formulations were administered as single oral dose and blood was sampled over 48 h. Metronidazole plasma concentrations were determined by a liquid chromatography mass spectrometry (LC-MS/MS) method. The plasma concentration vs. time profile was generated for each volunteer and the pharmacokinetic parameters Cmax, Tmax, AUC0-t, AUC0-∞, ke, and t1/2 were calculated using a noncompartmental model. Bioequivalence between pharmaceutical formulations was determined by calculating 90% CIs (Confidence Intervall) for the ratios of Cmax, AUC0-t, and AUC0-∞ values for test and reference using log-transformed data. 22 healthy volunteers (11 men, 11 women; mean (SD) age, 28 (6.5) years [range, 21-45 years]; mean (SD) weight, 66 (9.3) kg [range, 51-81 kg]; mean (SD) height, 169 (6.5) cm [range, 156-186 cm]) were enrolled in and completed the study. The 90% CIs for Cmax (0.92-1.06), AUC0-t (0.97-1.02), and AUC0-∞ (0.97-1.03) values for the test and reference products fitted in the interval of 0.80-1.25 proposed by most regulatory agencies, including the Brazilian agency ANVISA. No clinically significant adverse effects were reported. After pharmacokinetics analysis, it concluded that test 250 mg metronidazole formulation is bioequivalent to the reference product according to the Brazilian agency requirements.

Self-monitoring of Blood Glucose: One STeP Forward?

In times of short health care budgets, reimbursement for self-monitoring of blood glucose (SMBG) in diabetes patients without insulin treatment is subject to debate. The Structured Testing Program (STeP) trial found a positive correlation of test frequency and improved hemoglobin A1c (HbA1c) levels in poorly controlled type 2 diabetes patients not treated with insulin.

Elevated γ-glutamyltransferase

Reply To: "T Regulatory Cell Number and Function: The Autoimmune Traits in Liver Diseases"

Quantitative Accuracy of Attenuation Correction in the Philips Ingenuity TF Whole-body PET/MR System: a Direct Comparison with Transmission-based Attenuation Correction

OBJECTIVE: Evaluation of the quantitative accuracy of MR-based attenuation correction (MRAC) in the Philips Ingenuity TF whole-body PET/MR. MATERIALS AND METHODS: In 13 patients, PET emission data from the PET/MR were reconstructed using two different methods for attenuation correction. In the first reconstruction, the vendor-provided standard MRAC was used. In the second reconstruction, a coregistered transmission-based attenuation map from a second immediately preceding investigation with a stand-alone Siemens ECAT EXACT HR(+) PET scanner was used (TRAC). The two attenuation maps were compared regarding occurrence of segmentation artifacts in the MRAC procedure. Standard uptake values (SUVs) of multiple VOIs (liver, cerebellum, hot focal structures at various locations in the trunk) were compared between both reconstructed data sets. Furthermore, a voxel-wise intensity correlation analysis of both data sets in the lung and trunk was performed. RESULTS: VOI averaged SUV differences between MRAC and TRAC were as follows (relative differences, mean ± standard deviation): (+12 ± 6) % cerebellum, (-4 ± 9) % liver, (-2 ± 11) % hot focal structures. The fitted slopes of the voxel-wise correlations in the lung and trunk were 0.87 ± 0.17 and 0.95 ± 0.10 with averaged adjusted R (2) values of 0.96 and 0.98, respectively. These figures include two instances with partially erroneous lung segmentation due to artifacts in the underlying MR images. CONCLUSION: The MR-based attenuation correction implemented on the Philips Ingenuity PET/MR provides reasonable quantitative accuracy. On average, deviations from TRAC-based results are small (on the order of 10 %  or below) across the trunk, but due to interindividual variability of the segmentation quality, deviations of more than 20 %  can occur. Future improvement of the segmentation quality would help to increase the quantitation accuracy further and to reduce the inter-subject variability.

Femtomolar Inhibitors Bind to 5'-Methylthioadenosine Nucleosidases with Favorable Enthalpy and Entropy

5'-Methylthioadenosine/S-adenosylhomocysteine nucleosidase (MTAN) catalyzes the hydrolytic cleavage of adenine from methylthioadenosine (MTA). Inhibitor design and synthesis informed by transition state analysis have developed femtomolar inhibitors for MTANs, among the most powerful known noncovalent enzyme inhibitors. Thermodynamic analyses of the inhibitor binding reveals a combination of highly favorable contributions from enthalpic (-24.7 to -4.0 kcal mol(-1)) and entropic (-10.0 to 6.4 kcal mol(-1)) interactions. Inhibitor binding to similar MTANs from different bacterial species gave distinct energetic contributions from similar catalytic sites. Thus, binding of four transition state analogues to EcMTAN and SeMTAN is driven primarily by enthalpy, while binding to VcMTAN is driven primarily by entropy. Human MTA phosphorylase (hMTAP) has a transition state structure closely related to that of the bacterial MTANs, and it binds tightly to some of the same transition state analogues. However, the thermodynamic signature of binding of an inhibitor to hMTAP differs completely from that with MTANs. We conclude that factors other than first-sphere catalytic residue contacts contribute to binding of inhibitors because the thermodynamic signature differs between bacterial species of the same enzyme.

BRAF Mutation, NRAS Mutation, and the Absence of an Immune-Related Expressed Gene Profile Predict Poor Outcome in Patients with Stage III Melanoma

Prediction of outcome for melanoma patients with surgically resected macroscopic nodal metastases is very imprecise. We performed a comprehensive clinico-pathologic assessment of fresh-frozen macroscopic nodal metastases and the preceding primary melanoma, somatic mutation profiling, and gene expression profiling to identify determinants of outcome in 79 melanoma patients. In addition to disease stage 4 years, 90% confidence interval): the presence of a nodular component in the primary melanoma (6.8, 0.6-76.0), and small cell size (11.1, 0.8-100.0) or low pigmentation (3.0, 0.8-100.0) in the nodal metastases. Absence of BRAF mutation (20.0, 1.0-1000.0) or NRAS mutation (16.7, 0.6-1000.0) were both favorable prognostic factors. A 46-gene expression signature with strong overrepresentation of immune response genes was predictive of better survival (10.9, 0.4-325.6); in the full cohort, median survival was >100 months in those with the signature, but 10 months in those without. This relationship was validated in two previously published independent stage III melanoma data sets. We conclude that the presence of BRAF mutation, NRAS mutation, and the absence of an immune-related expressed gene profile predict poor outcome in melanoma patients with macroscopic stage III disease.Journal of Investigative Dermatology advance online publication, 30 August 2012; doi:10.1038/jid.2012.283.

[Satisfied General Practitioners and Critical Nursing Staff - Problems of Interprofessional Cooperation in the Home Care of Dementia Patients.]

Analysis of views of general practioners and nurses of interprofessional cooperation between general practititoners and nurses in the ambulatory care of dementia patients is presented.A survey was carried out among general practitioners and nurses caring for community dwelling dementia patients in Hamburg.The majority of GPs and nurses consider interprofessional cooperation to be good and beneficial for their own work. GPs are generally more positive about the quality of cooperation than nurses. Joint sessions for planning and evaluation of care are seldom. Even so, more GPs than nurses evaluate the frequency of these meetings to be sufficient. Although nurses are more critical about the quality of the cooperation with the GPs, they seldom address the GP to express their criticism.To make cooperation work, the matter should be part of the training of both physicians and nurses and the hierarchy between the 2 groups should be reduced.

Sialidase, Chondroitinase ABC and Combination Therapy After Spinal Cord Contusion Injury

Axon regeneration in the central nervous system is severely hampered, limiting functional recovery. This is due, in part, to endogenous axon regeneration inhibitors that accumulate at the injury site. Therapeutic targeting of these inhibitors and their receptors may facilitate axon outgrowth and enhance recovery. A rat model of spinal cord contusion injury was used to test the effects of two bacterial enzyme therapies that target independent axon regeneration inhibitors, sialidase (V. cholerae) and chondroitinase ABC (ChABC, P. vulgaris). The two enzymes, individually and in combination, were infused for two weeks via implanted osmotic pumps to the site of a moderate thoracic spinal cord contusion injury. Sialidase was completely stable whereas ChABC retained >30% of its activity in vivo over the two-week infusion period. Immunohistochemistry revealed that infused sialidase acted robustly throughout the spinal cord grey and white matter, whereas ChABC activity was more intense superficially. Sialidase treatment alone resulted in improved behavioral and anatomical outcomes. Rats treated exclusively with sialidase showed significantly increased hindlimb motor function, evidenced by higher BBB and BBB subscores, and fewer stepping errors on a horizontal ladder. Sialidase-treated rats also had increased serotonergic axons caudal to the injury. ChABC treatment, in contrast, did not enhance functional recovery or alter axon numbers after moderate spinal cord contusion injury, and damped the response of sialidase in the dual enzyme treatment group. We conclude that sialidase infusion enhanced recovery from spinal cord contusion injury, and that combining sialidase with ChABC failed to improve outcomes. Key words: axon regeneration; ganglioside; chondroitin sulfate; motor behavior; serotonergic axons.

The Influence of Preoperative Serum Anticholinergic Activity and Other Risk Factors for the Development of Postoperative Cognitive Dysfunction After Cardiac Surgery

OBJECTIVE: Patients undergoing cardiac surgery are at risk for postoperative cognitive dysfunction (POCD). Evaluating preoperative risk factors represents 1 possible way to minimize the postoperative risk of cognitive dysfunction. We investigated marked deterioration in cholinergic neurotransmission as 1 such potential risk factor for transient and long-lasting POCD. Serum anticholinergic activity (SAA) has already been described as a risk factor for developing delirium in an elderly study population. However, the role of SAA for long-lasting POCD is unknown. METHODS: Following local ethics board approval and written informed consent, we recruited a cohort of patients aged ≥ 55 years undergoing cardiac surgery. Before surgery, levels of SAA were measured and a battery of neuropsychologic tests (NPTs) was applied. S100 calcium binding protein ß concentration was measured intraoperatively. Pre-, intra-, and postoperative patient-specific characteristics were recorded. The NPTs were repeated 3 months after hospital discharge to evaluate 3-month POCD. A group of nonsurgical patients (n = 34) was recruited as control subjects to adjust NPT scores, using reliable methods for the change index. Logistic multivariate regression was used to evaluate independent predictors of POCD. RESULTS: One hundred fifty-four patients were screened before surgery, and 117 completed the second NPT. POCD was identified in 25.6% of patients. In contrast to intraoperatively increased S100 calcium binding protein ß, preoperative SAA was not associated with POCD following adjustment for covariates. CONCLUSIONS: Preoperatively increased SAA did not predict POCD 3 months after cardiac surgery.

Evolution and Analysis of Genetic Networks for Stable Cellular Growth and Regeneration

A computational model is presented that simulates stable growth of cellular structures that are in some cases capable of regeneration. In the model, cellular growth is governed by a gene regulatory network. By evolving the parameters and structure of the genetic network using a modified evolution strategy, a dynamically stable state can be achieved in the developmental process, where cell proliferation and cell apoptosis reach an equilibrium. The results of evolution with different setups in fitness evaluation during the development are compared with respect to their regeneration capability as well as their gene regulatory network structure. Network motifs responsible for stable growth and regeneration that emerged from the evolution are also analyzed. We expect that our findings can help to gain a better understanding of the process of growth and regeneration inspired by biological systems, in order to solve complex engineering problems, such as the design of self-healing materials.

Surgery in Adults: Temporal Resections

Surgical Management of Medial Extratemporal Epilepsy

Pediatric Functional Hemispherectomy: Outcome in 92 Patients

The revival of epilepsy surgery after the introduction of modern presurgical evaluation procedures has led to an increase in hemispherectomy or hemispherotomy procedures. Since a large part of our pediatric series was done using a newer hemispherotomy technique, we focus mainly on the outcomes after a recently developed hemispherotomy technique (transsylvian keyhole).

Diagnostic Performance of Whole Brain Volume Perfusion CT in Intra-axial Brain Tumors: Preoperative Classification Accuracy and Histopathologic Correlation

To evaluate the preoperative diagnostic power and classification accuracy of perfusion parameters derived from whole brain volume perfusion CT (VPCT) in patients with cerebral tumors.

Long-term Outcome of an Unusual Haemophilic Pseudotumour

Haemophilia, a lifelong congenital bleeding disease, is a highly demanding disorder, due to the costs of its replacement therapy. In the absence of this pivotal treatment, life expectancy and quality of life are deleteriously affected. As illustration, we present a 14 years long follow-up of a patient with severe haemophilia A, treated sporadically with fresh plasma, cryoprecipitate and factor concentrates, who developed a giant iliopsoas pseudotumor. Since he was an infant, under on demand therapy with fresh frozen plasma, cryoprecipitate and low doses of factor concentrates he presented many spontaneous bleedings, developing multiple disabling arthropathies. At the age of 14 years, an iliopsoas hematoma occurred, which relapsed several times, developing an iliopsoas pseudotumour. After 5 years, sepsis with Klebsiella was diagnosed. A CT scan revealed fistula between the pseudotumor and the gut. Under antibiotics, the evolution of sepsis improved, but over a period of 10 months 5 episodes of haematemesis and melena, followed by one episode of macroscopic haematuria occurred; two months later he developed an inguino-crural mass, which fistulized through the abdominal wall. A mixt german-romanian team solved the clinical concern. After 108 hospitalization days and consumption of 104840 IU factor VIII he left the clinic in good condition. One year later, the temporary colostomy with anus praeter was closed. The follow-up reveals now, after almost 10 years with favourable outcome, that the patient is well, active within his family and profession.

[Morbidity and Mortality of Patients with Haemophilia in Germany--update 2010/2011]

Since 1982 an annual multicentric survey regarding the epidemiology of patients suffering of haemophilia is performed with support of haemophilia treating centres of any size. Again, the actual compilation is resting upon a broad database returning to over 30 years of inquiry well representing both the actual and retrospective status of mortality. Prompted was exclusively information about patients with haemophilia A, B and von Willebrand disease. In particular anonymous data concerning the preceeding 12 months about number of treated patients, type and severity of illness, HIV-status and detailed information about causes of death was inquired. This data was merged with existing data and analysed statistically. In the 2009/2010 survey, a total number of 9448 patients with bleeding disorders have been reported from 47 participating centres. Despite mortality from HIV in patients with haemophilia is keeping on decreasing, HIV still remains an important factor as an HIV/HCV coinfection seems to increase risk of progression of severe liver disease. In the actual investigation the findings of the foundation "Humanitäre Hilfe für durch Blutprodukte HIV-infizierte Personen" were compared for the first time to our data. Time trends were visualised with a moving average. Our investigation has a smaller number of deceased patients, but contains consistent data.

Antegrade Flow Across Incomplete Vessel Occlusions Can Be Distinguished from Retrograde Collateral Flow Using 4-dimensional Computed Tomographic Angiography

In acute stroke patients with intracranial vessel occlusion, angiographic demonstration of antegrade contrast opacification distal to the occlusion site (termed the "clot outline sign") has been associated with higher rates of vessel recanalization. We sought to determine whether antegrade flow can be demonstrated on timeresolved 4-dimensional computed tomographic angiography (4-dimensional CTA), whether it can be distinguished from retrograde collateral flow, and if it can be used to predict early recanalization.

Evaluation of Organochlorine Pesticides in Breast Milk Samples in Turkey Applying Features of the Partial Order Technique

Exposure data to organochlorine pesticides (OCPs) of mothers' breast milk samples were measured in different locations in the Taurus Mountains/Turkey. Villages at five altitudes were looked upon. The data analysis method applied is the Hasse diagram technique, based on Discrete Mathematics. For each of the five regions, one Hasse diagram was calculated and showed the same maximal object, namely, p-p'-Dichlorodiphenyldichloroethane (DDE). In a further data investigation step, we performed the similarity analyses of the locations in order to quantify the similarities and incomparabilities. It was revealed that no correlation exists between most altitudes and the pattern of contamination of breast milk samples. The two lowest situated areas, Sahil at sea level and Ciftlikköy at 121 m above sea level were similar. We reduced the datasets by applying the interval analysis followed by similarity analyses. We observed that endrin (END1), mirex (MIRE), and pentachlorobenzene (PECB) reduce most severely the similarity.

Schistosome-derived Omega-1 Drives Th2 Polarization by Suppressing Protein Synthesis Following Internalization by the Mannose Receptor

Omega-1, a glycosylated T2 ribonuclease (RNase) secreted by Schistosoma mansoni eggs and abundantly present in soluble egg antigen, has recently been shown to condition dendritic cells (DCs) to prime Th2 responses. However, the molecular mechanisms underlying this effect remain unknown. We show in this study by site-directed mutagenesis of omega-1 that both the glycosylation and the RNase activity are essential to condition DCs for Th2 polarization. Mechanistically, we demonstrate that omega-1 is bound and internalized via its glycans by the mannose receptor (MR) and subsequently impairs protein synthesis by degrading both ribosomal and messenger RNA. These experiments reveal an unrecognized pathway involving MR and interference with protein synthesis that conditions DCs for Th2 priming.

PET/MR for Therapy Response Evaluation in Malignant Lymphoma: Initial Experience

OBJECT: To evaluate the feasibility of positron emission tomography/magnetic resonance imaging (PET/MR) with (18)fluoro-2-deoxyglucose (FDG) for therapy response evaluation of malignant lymphoma. MATERIALS AND METHODS: Nine patients with malignant lymphoma who underwent FDG-PET/MR before and after chemotherapy were included in this retrospective study. Average time between the two scans was 70 days. The scans were evaluated independently by two nuclear medicine physicians. The Ann Arbor classification was used to describe lymphoma stage. Furthermore, the readers also rated PET image quality using a five point scale. Weighted kappa (κ) was used to calculate interrater agreement. RESULTS: The initial scan showed foci of increased FDG uptake in all patients, with Ann Arbor stage varying between I and IV. In the follow-up examination, all but one patient showed complete response to chemotherapy. PET image quality was rated as very good or excellent for all scans. Interrater agreement was excellent regarding Ann Arbor stage (κ = 0.97) and good regarding image quality (κ = 0.41). CONCLUSION: PET/MR shows promising initial results for therapy response evaluation in lymphoma patients.

Effect of Continuous Venovenous Hemofiltration Dose on Achievement of Adequate Vancomycin Trough Concentrations

The vancomycin dose necessary for the achievement of target serum trough concentrations during continuous venovenous hemofiltration (CVVH) remains to be elucidated. This was a retrospective cohort study of critically ill adults at a tertiary medical center on concurrent CVVH and vancomycin between 2006 and 2010 with a steady-state vancomycin trough concentration. The 87 included patients were grouped according to low (≤30 ml/kg/h; n = 10) or high (>30 ml/kg/h; n = 77) CVVH hemofiltration rate (HFR) for analysis. Vancomycin goal trough achievement occurred in only 32 (37%) patients. The primary endpoint of trough attainment significantly differed between HFR subgroups: 90% versus 30% in low- and high-HFR individuals, respectively (P < 0.001). Patients with subtherapeutic trough concentrations had a median (interquartile range) HFR of 40 ml/kg/h (range, 37 to 47 ml/kg/h) compared to 36 ml/kg/h (range, 30 to 39 ml/kg/h) in those who achieved the trough goal. Irrespective of goal trough, an inverse correlation existed between HFR and serum vancomycin concentration (r = -0.423; P < 0.001). In the subgroup of 14 methicillin-resistant Staphylococcus aureus (MRSA) patients, trough achievement was similar to the aggregate cohort (36%). Mortality at 28 days was unrelated to trough achievement in both the overall sample (P = 0.516) and in culture-positive MRSA patients (P = 0.396). Critically ill patients undergoing CVVH therapy may experience clinically significant reductions in goal vancomycin troughs. The results of the present study justify prospective evaluations in this population to determine the optimal vancomycin dosing strategy for attainment of goal trough concentrations.

Inhibition of Inflammatory CD4 T Cell Activity by Murine Liver Sinusoidal Endothelial Cells

BACKGROUND & AIMS: The liver can mitigate the inflammatory activity of infiltrating T cells by mechanisms that are not entirely clear. Here we investigated the role of liver sinusoidal endothelial cells (LSECs) in regulating the activity of inflammatory CD4 T cells. METHODS: Interactions between T helper (Th) 1 or Th17 cells and LSEC were studied by intravital microscopy and by in vitro stimulation assays. RESULTS: Circulating CD4 T cells established lasting and repeated interactions with liver endothelium in vivo. Stimulation of Th1 and Th17 cells by LSEC greatly inhibited their capacity to secrete interferon-γ or interleukin-17 in vitro; in contrast, stimulation by dendritic cells (DCs) resulted in considerable secretion of both cytokines. Cytokine release by Th1 or Th17 cells seemed to be actively suppressed by LSEC, as indicated by the inhibition of cytokine secretion even in the presence of Th1- and Th17-promoting DC. This inhibition of CD4 T cell effector function seemed to depend on the dominance of inhibitory over activating co-stimulatory signals on LSEC, since (1) cytokine secretion could be restored by increased CD28 co-activation; (2) LSEC from interleukin-10(-/-) mice, which manifest increased activating signals, such as MHC II, and decreased inhibitory signals, such as PD-L1, failed to suppress cytokine secretion; and (3) cytokine secretion by Th1 or Th17 cells that lacked PD-1, the ligand for inhibitory PD-L1, could not be suppressed by LSEC. CONCLUSIONS: LSEC inhibit inflammatory cytokine secretion of Th1 and Th17 effector CD4 T cells in dependence of interleukin-10 and PD-1.

[Focal Neuropathy]

The differential diagnosis of focal neuropathy continues to be one of the main tasks in peripheral neurology and clinical neurophysiology. In this respect a detailed medical history a topologically oriented clinical examination based on profound anatomical knowledge and a targeted neurophysiological examination are the cornerstones of successful diagnosis. Refined imaging techniques, such as sonography and magnetic resonance imaging provide - additional and valuable morphological information especially in atypical or unresolved cases. The aim of this overview is therefore to summarize the spectrum of focal neuropathy and diagnostic approaches, particularly in terms of a well-founded differential diagnosis.

Nanolesions Induced by Heavy Ions in Human Tissues: Experimental and Theoretical Studies

The biological effects of energetic heavy ions are attracting increasing interest for their applications in cancer therapy and protection against space radiation. The cascade of events leading to cell death or late effects starts from stochastic energy deposition on the nanometer scale and the corresponding lesions in biological molecules, primarily DNA. We have developed experimental techniques to visualize DNA nanolesions induced by heavy ions. Nanolesions appear in cells as "streaks" which can be visualized by using different DNA repair markers. We have studied the kinetics of repair of these "streaks" also with respect to the chromatin conformation. Initial steps in the modeling of the energy deposition patterns at the micrometer and nanometer scale were made with MCHIT and TRAX models, respectively.

Broadband, Diode Pumped Yb:SiO2 Multicomponent Glass Laser

Fabrication, spectroscopic properties, and laser performance of a Yb:SiO(2) multicomponent glass have been investigated in this paper. The glass system composed of SiO(2), Al(2)O(3), and La(2)O(3) excels in terms of a high thermal stress resistance compared to other laser glasses. The laser experiments were conducted with a 3.4 mm thick and 0.9 mol. % Y(2)O(3) doped sample. A maximum slope efficiency of 51%, a maximum optical to optical efficiency of 42%, and a tuning range from 1010-1090 nm was realized. Due to the promising laser properties and a straightforward fabrication technique it may well qualify as an alternative gain medium in high-energy, ultrashort pulse laser systems.

Development of a Nondestructive Leak Testing Method Utilizing the Head Space Analyzer for Ampoule Products Containing Ethanol-based Solutions

The application of a head space analyzer for oxygen concentration was examined to develop a novel ampoule leak test method. Studies using ampoules filled with ethanol-based solution and with nitrogen in the headspace demonstrated that the head space analysis (HSA) method showed sufficient sensitivity in detecting an ampoule crack. The proposed method is the use of HSA in conjunction with the pretreatment of an overpressurising process known as bombing to facilitate the oxygen flow through the crack in the ampoule. The method was examined in comparative studies with a conventional dye ingress method, and the results showed that the HSA method exhibits sensitivity superior to the dye method. The results indicate that the HSA method in combination with the bombing treatment provides potential application as a leak test for the detection of container defects not only for ampoule products with ethanol-based solutions, but also for testing lyophilized products in vials with nitrogen in the head space. LAY ABSTRACT: The application of a head space analyzer for oxygen concentration was examined to develop a novel ampoule leak test method. The proposed method is the use of head space analysis (HSA) in conjunction with the pretreatment of an overpressurising process known as bombing to facilitate oxygen flow through the crack in the ampoule for use in routine production. The result of the comparative study with a conventional dye leak test method indicates that the HSA method in combination with the bombing treatment can be used as a leak test method, enabling detection of container defects.

Impaired Cerebrovascular Autoregulation in Patients with Severe Sepsis and Sepsis-associated Delirium

ABSTRACT: INTRODUCTION: Sepsis-associated delirium (SAD) increases morbidity in septic patients and, therefore, factors contributing to SAD should be further characterized. One possible mechanism might be the impairment of cerebrovascular autoregulation (AR) by sepsis, leading to cerebral hypo- or hyperperfusion in these haemodynamically unstable patients. Therefore, the present study investigates the relationship between the incidence of SAD and the status of AR during sepsis. METHODS: Cerebral blood flow velocity was measured using transcranial Doppler sonography and was correlated with the invasive arterial blood pressure curve to calculate the index of AR Mx (Mx>0.3 indicates impaired AR). Mx was measured daily during the first 4 days of sepsis. Diagnosis of a SAD was performed using the confusion assessment method for ICU (CAM-ICU) and, furthermore the predominant brain electrical activity in electroencephalogram (EEG) both at day 4 after reduction of sedation to RASS >-2. RESULTS: 30 critically ill adult patients with severe sepsis or septic shock (APACHE II 32 ± 6) were included. AR was impaired at day 1 in 60%, day 2 in 59%, day 3 in 41% and day 4 in 46% of patients; SAD detected by CAM-ICU was present in 76 % of patients. Impaired AR at day 1 was associated with the incidence of SAD at day 4 (p = 0.035). CONCLUSIONS: AR is impaired in the great majority of patients with severe sepsis during the first two days. Impaired AR is associated with SAD, suggesting that dysfunction of AR is one of the trigger mechanisms contributing to the development of SAD. TRIAL REGISTRATION: clinicalTrials.gov ID NCT01029080.

High-energy, Ceramic-disk Yb:LuAG Laser Amplifier

We report the first short-pulse amplification results to several hundred millijoule energies in ceramic Yb:LuAG. We have demonstrated ns-pulse output from a diode-pumped Yb:LuAG amplifier at a maximum energy of 580 mJ and a peak optical-to-optical efficiency of 28% at 550 mJ. In cavity dumped operation of a nanosecond oscillator we obtained 1 mJ at up to 100 Hz repetition rate. A gain bandwidth of 5.4 nm was achieved at room temperature by measuring the small-signal single-pass gain. Furthermore, we compared our results with Yb:YAG within the same amplifier system.

Higher Nitrate-reducer Diversity in Macrophyte-colonized Compared to Unvegetated Freshwater Sediment

Freshwater macrophytes stimulate rhizosphere-associated coupled nitrification-denitrification and are therefore likely to influence the community composition and abundance of rhizosphere-associated denitrifiers and nitrate reducers. Using the narG gene, which encodes the catalytic subunit of the membrane-bound nitrate reductase, as a molecular marker, the community composition and relative abundance of nitrate-reducing bacteria were compared in the rhizosphere of the freshwater macrophyte species Littorella uniflora and Myriophyllum alterniflorum to nitrate-reducing communities in unvegetated sediment. Microsensor analysis indicated a higher availability of oxygen in the rhizosphere compared to unvegetated sediment, with a stronger release of oxygen from the roots of L. uniflora compared to M. alterniflorum. Comparison of narG clone libraries between samples revealed a higher diversity of narG phylotypes in association with the macrophyte rhizospheres compared to unvegetated sediment. Quantitative PCR targeting narG- and 16S rRNA-encoding genes pointed to a selective enrichment of narG gene copies in the rhizosphere. The results suggested that the microenvironment of macrophyte rhizospheres, characterized by the release of oxygen and labile organic carbon from the root system, had a stimulating effect on the diversity and relative abundance of rhizosphere-associated nitrate reducers.

LIN28B Induces Neuroblastoma and Enhances MYCN Levels Via Let-7 Suppression

LIN28B regulates developmental processes by modulating microRNAs (miRNAs) of the let-7 family. A role for LIN28B in cancer has been proposed but has not been established in vivo. Here, we report that LIN28B showed genomic aberrations and extensive overexpression in high-risk neuroblastoma compared to several other tumor entities and normal tissues. High LIN28B expression was an independent risk factor for adverse outcome in neuroblastoma. LIN28B signaled through repression of the let-7 miRNAs and consequently resulted in elevated MYCN protein expression in neuroblastoma cells. LIN28B-let-7-MYCN signaling blocked differentiation of normal neuroblasts and neuroblastoma cells. These findings were fully recapitulated in a mouse model in which LIN28B expression in the sympathetic adrenergic lineage induced development of neuroblastomas marked by low let-7 miRNA levels and high MYCN protein expression. Interference with this pathway might offer therapeutic perspectives.

Exon-level Expression Analyses Identify MYCN and NTRK1 As Major Determinants of Alternative Exon Usage and Robustly Predict Primary Neuroblastoma Outcome

Using mRNA expression-derived signatures as predictors of individual patient outcome has been a goal ever since the introduction of microarrays. Here, we addressed whether analyses of tumour mRNA at the exon level can improve on the predictive power and classification accuracy of gene-based expression profiles using neuroblastoma as a model.

Methylthioadenosine Deaminase in an Alternative Quorum Sensing Pathway in Pseudomonas Aeruginosa

Pseudomonas aeruginosa possesses an unusual pathway for 5'-methylthioadenosine (MTA) metabolism involving deamination to 5'-methylthioinosine (MTI) followed by N-ribosyl phosphorolysis to hypoxanthine and 5-methylthio-α-d-ribose 1-phosphate. The specific MTI phosphorylase of P. aeruginosa has been reported [Guan, R., Ho, M. C., Almo, S. C., and Schramm, V. L. (2011) Biochemistry 50, 1247-1254], and here we characterize MTA deaminase from P. aeruginosa (PaMTADA). Genomic analysis indicated the PA3170 locus to be a candidate for MTA deaminase (MTADA). Protein encoded by PA3170 was expressed and shown to deaminate MTA with 40-fold greater catalytic efficiency for MTA than for adenosine. The k(cat)/K(m) value of 1.6 × 10(7) M(-1) s(-1) for MTA is the highest catalytic efficiency known for an MTA deaminase. 5'-Methylthiocoformycin (MTCF) is a 4.8 pM transition state analogue for PaMTADA but causes no significant inhibition of human adenosine deaminase or MTA phosphorylase. MTCF is permeable to P. aeruginosa and exhibits an IC(50) of 3 nM on cellular PaMTADA activity. PaMTADA is the only activity in P. aeruginosa extracts to act on MTA. MTA and 5-methylthio-α-d-ribose are involved in quorum sensing pathways; thus, PaMTADA is a potential target for quorum sensing. The crystal structure of PaMTADA in complex with MTCF shows the transition state mimic 8(R)-hydroxyl group in contact with a catalytic site Zn(2+), the 5'-methylthio group in a hydrophobic pocket, and the transition state mimic of the diazepine ring in contact with a catalytic site Glu.

Lambert-Eaton Myasthenic Syndrome Presenting with a "dropped Head Syndrome" and Associated with Antibodies Against N-type Calcium Channels

Trends in Presurgical Evaluation and Surgical Treatment of Epilepsy at One Centre from 1988-2009

BACKGROUND: Presurgical epilepsy diagnostics and surgical treatment have become standard procedures of neurology. Here, we report on presurgical patients grouped according to their underlying pathology by giving results of presurgical evaluation, surgical therapy and long-term follow-up between 1989-2009 and describe trends over this period. PATIENTS AND METHODS: Data of prospectively documented presurgical patients were retrospectively analysed. Trends were evaluated by a year-by-year analysis. RESULTS: 2684 presurgical patients underwent presurgical work-up, 1721 of whom (64.1%) went on to resective surgery. The largest presurgical/surgical group was mediotemporal lobe epilepsy with hippocampal sclerosis (29.5%/35.4%). Of all operated patients, 1160 (67.4%) had a follow-up of ≥2 years. A total of 586 were continuously seizure-free without auras (50.5%; benign tumours: 61.0%; focal cortical dysplasia: 57.6%; mediotemporal lobe epilepsy with hippocampal sclerosis: 49.4%; non-lesional: 27.6%). Based on the number of the presurgically studied patients, we calculated as a novel measure of the effect of a presurgical/surgical programme an 'intention-to treat seizure-freedom' rate of 32.4%. Over time, the number of patients undergoing evaluation, but also of those not suitable or agreeable for invasive measures increased. Annual numbers of resective procedures remained stable. Average epilepsy duration of patients admitted for presurgical assessment increased. The proportion of patients with benign tumours declined. Intracranial studies and MRI-histopathology discrepancies decreased. Seizure-freedom rates remained constant. CONCLUSIONS: Epilepsy surgery is highly effective, especially in patients with clearly defined focal pathologies. At this specialised centre, there is a trend towards growing numbers of difficult patients and increasing epilepsy duration prior to referral for presurgical assessment.

Leveraging Ethnic Group Incidence Variation to Investigate Genetic Susceptibility to Glioma: a Novel Candidate SNP Approach

Objectives: Using a novel candidate SNP approach, we aimed to identify a possible genetic basis for the higher glioma incidence in Whites relative to East Asians and African-Americans. Methods:  We hypothesized that genetic regions containing SNPs with extreme differences in allele frequencies across ethnicities are most likely to harbor susceptibility variants. We used International HapMap Project data to identify 3,961 candidate SNPs with the largest allele frequency differences in Whites compared to East Asians and Africans and tested these SNPs for association with glioma risk in a set of White cases and controls. Top SNPs identified in the discovery dataset were tested for association with glioma in five independent replication datasets. Results: No SNP achieved statistical significance in either the discovery or replication datasets after accounting for multiple testing or conducting meta-analysis. However, the most strongly associated SNP, rs879471, was found to be in linkage disequilibrium with a previously identified risk SNP, rs6010620, in RTEL1. We estimate rs6010620 to account for a glioma incidence rate ratio of 1.34 for Whites relative to East Asians. Conclusion: We explored genetic susceptibility to glioma using a novel candidate SNP method which may be applicable to other diseases with appropriate epidemiologic patterns.

Comparative Investigation in Stray Light Measurement in the Human Eye

Identifying Transcriptional MiRNA Biomarkers by Integrating High-throughput Sequencing and Real-time PCR Data

Using both high-throughput sequencing and real-time PCR, the miRNA transcriptome can be analyzed in complementary ways. We describe the necessary bioinformatics pipeline, including software tools, and key methodological steps in the process, such as adapter removal, read mapping, normalization, and multiple testing issues for biomarker identification. The methods are exemplified by the analysis of five favorable (event-free survival) vs. five unfavorable (died of disease) neuroblastoma tumor samples with a total of over 188 million reads.

Filamentous Bacteria Transport Electrons over Centimetre Distances

Oxygen consumption in marine sediments is often coupled to the oxidation of sulphide generated by degradation of organic matter in deeper, oxygen-free layers. Geochemical observations have shown that this coupling can be mediated by electric currents carried by unidentified electron transporters across centimetre-wide zones. Here we present evidence that the native conductors are long, filamentous bacteria. They abounded in sediment zones with electric currents and along their length they contained strings with distinct properties in accordance with a function as electron transporters. Living, electrical cables add a new dimension to the understanding of interactions in nature and may find use in technology development.

Seasonal Methane Oxidation Potential in Manure Crusts

Organic crusts on liquid manure storage tanks harbor ammonia- and nitrite-resistant methane oxidizers and may significantly reduce methane emissions. Methane oxidation potential (0.6 mol CH(4) m(-2) d(-1)) peaked during fall and winter, after 4 months of crust development. Consequences for methane mitigation potential of crusts are discussed.

[42-year-old Woman with Pain of Wrist After a Fall]

Activation of Thalamus in Motor Imagery Results from Gating by Hypnosis

The ability to mentally imagine the performance of automatic movements has been well-established being employed in sports and physiotherapy as a tool for motor learning and rehabilitation. This is probably mediated by engagement of the same brain areas as during real motor performance. Here we investigated the effect of hypnotic trance on the cerebral activation pattern engaged in motor imagery in 16 healthy, right-handed subjects using fMRI. Motor imagery as compared with rest was related to activations in the left medial frontal areas (preSMA/SMA), prefrontal- and frontal areas, putamen and inferior parietal areas. When compared with performance of the same movements motor imagery resulted in activation of the left middle frontal cortex, precuneus, and posterior cingulate. Under hypnotic trance there was one extra-activation in the left thalamus which occurred specifically in the motor imagery condition. The regional beta indices were highly correlated among the areas of the cortical-subcortical motor network. Our data accord with the notion that hypnotic trance enhances the motor control circuit engaged in motor imagery by modulating the gating function of the thalamus.

Embedding Resorcinarene Cavitands in Lipid Vesicles()

A fluorescently labeled resorcinarene cavitand has been successfully embedded in DLPC lipid vesicles and imaged using confocal microscopy. The cavitand resides exclusively in the bilayer.

[Updated Quality Requirements Regarding Secondary Differentiated Ultrasound Examination in Prenatal Diagnostics (= DEGUM Level II) in the Period from 18 + 0 to 21 + 6 Weeks of Gestation]

[The Morality of Nanotechnology]

Nanotechnology is a set of knowledge, techniques, and practices in studying and exploring new properties of materials that arise when manipulated at the atomic and molecular levels. The technical possibility of organizing and controlling matter at the smallest dimensions and units can result in profound changes in industrial production processes and have significant moral impacts on human relations, organization of the current social order, and even life as a phenomenon. However, moral reflection on nanotechnology has been criticized over the assertion that nanotechnology fails to raise any new ethical issue, for example. The current article discusses the limits of this claim by presenting two aspects that distinguish between nanotechnology and earlier biotechnoscientific advances in terms of their ethical implications: (a) uncertainty as an epistemic characteristic and (b) the threat to the current symbolic character of DNA as the "code of life".

Amniotic Bands As a Cause of Congenital Anterior Staphyloma

BACKGROUND: Congenital anterior staphyloma is a rare, complex malformation syndrome of the anterior segment. Only a few reports on associated systemic malformations have been published. We herein present a rare manifestation of congenital anterior staphyloma (CAS) combined with amniotic band disruption syndrome (ABS). PATIENT AND METHODS: Shortly after birth, a massive enlargement of the left eye was observed in a female child. Furthermore, an extensive bilateral congenital cleft lip and cleft alveolar ridge with oblique facial cleft extending into the left medial canthal region, coloboma(s) of the left eyelids, extensive adhesions between lids and eye bulb, as well as circumferential grooves, clubfeet, and terminal transverse defects in both hands and feet were present. Due to severe progression of eye bulb protrusion with thinning of the sclera, enucleation of the left eye was performed at the age of 3 years in order to prevent complications including perforation of the globe and with the aim of improving cosmetic aspects. RESULTS: Histopathological examination of the enucleated eye disclosed findings typical of congenital anterior staphyloma, including massive corneal staphylomatic deformation with superficial vascularization and elapsed corneoscleral margin, destruction of Bowman's layer, absence of Descemet's layer, corneal endothelium, and angle structures. The lens was only partially formed, and had mainly dissolved. The neural retina appeared normal. The optic nerve disc revealed a pronounced excavation. Facial clefts, lid colobomas, congenital constriction bands, and amputation of distal limbs match ABS. This malformation complex develops in early pregnancy, probably prior to 35 days post conception. CONCLUSION: This is the first report on an association of these two rare complex congenital malformations, congenital anterior staphyloma and amniotic band syndrome. The anterior staphyloma was unilateral, and related to facial clefts and lid coloboma in the area adjacent to the anterior staphyloma. Furthermore, the systemic deformities are clearly due to the amniotic bands, and the timing of the development of both complex malformations seems to be similar. All findings suggest that the presence of amniotic bands is a causative factor for all observed abnormalities including anterior staphyloma.

Influence of Intragenic CCL3 Haplotypes and CCL3L Copy Number in HIV-1 Infection in a Sub-Saharan African Population

Two CCL3 haplotypes (HapA1 and Hap-A3) and two polymorphic positions shared by the haplotypes (Hap-2SNP (single nucleotide polymorphism)) were investigated together with CCL3L copy number (CN), for their role in HIV-1 disease. Hap-A1 was associated with protection from in utero HIV-1 infection: exposed uninfected (EU) infants had higher representation of wild type (WT)/Hap-A1 than infected infants (excluding intrapartum (IP)-infected infants), which maintained significance post maternal Nevirapine (mNVP) and viral load (MVL) correction (P=0.04; odds ratio (OR)=0.33). Mother-infant pair analyses showed the protective effect of Hap-A1 is dependent on its presence in the infant. Hap-A3 was associated with increased IP transmission: WT/Hap-A3 was increased in IP-transmitting vs non-transmitting (NT) mothers, and remained significant post mNVP and MVL correction (P=0.02; OR=3.50). This deleterious effect of Hap-A3 seemed dependent on its presence in the mother. Hap-2SNP was associated with lower CD4 count in the NT mothers (P=0.03). CCL3 Hap-A1 was associated with high CCL3L CN in total (P=0.001) and EU infants (P=0.006); the effect was not additive, however, having either Hap-A1 or high CCL3L CN was more significantly (P=0.0008) associated with protection from in utero infection than Hap-A1 (P=0.028) or high CCL3L CN (P=0.002) alone. Linkage disequilibrium between Hap-A1 and high CCL3L CN appears unlikely given that a Nigerian population showed an opposite relationship.Genes and Immunity advance online publication, 15 November 2012; doi:10.1038/gene.2012.51.

Single Umbilical Incision Laparoscopic Cholecystectomy: Results of the Prospective Trial of the Coelio Club

To assess the cosmetic outcome after single umbilical incision laparoscopic cholecystectomies (SILC) performed by the surgeons of the Coelio Club.

Mold Attacks a Beating Heart

No abstract available.

Prevalence of Cone Beam Computed Tomography Imaging Findings According to the Clinical Stage of Bisphosphonate-related Osteonecrosis of the Jaw

Investigations of cone beam computed tomography (CBCT) for bisphosphonate-related osteonecrosis of the jaw (BRONJ) imaging are rare. The purpose of this study was to investigate the prevalence of typical radiological findings of BRONJ in CBCT.

Position of the Conus Medullaris in Fetuses with Skeletal Dysplasia

To examine whether reduced conus distance (CD) measurement may be a marker for impaired growth of the fetal trunk in fetuses with certain types of skeletal dysplasia.

Hydrocephalus, Agenesis of the Corpus Callosum, and Cleft Lip/palate Represent Frequent Associations in Fetuses with Peters' Plus Syndrome and B3GALTL Mutations. - Fetal PPS Phenotypes, Expanded by Dandy Walker Cyst and Encephalocele

OBJECTIVE: Fetal pathology aims to recognize syndromal patterns of anomalies for goal-directed mutation analyses, genetic counseling, and early prenatal diagnosis in consecutive pregnancies. Here, we report on five fetuses with Peters' plus syndrome (PPS) from two distinct families aborted after prenatal ultrasound diagnosis of hydrocephaly. METHOD: We performed fetal autopsies and molecular analyses. RESULTS: Among 44 fetuses with prenatally diagnosed hydrocephaly, four fetuses of 16 to 21 gestational weeks presented with additional cleft lip/palate and/or agenesis of the corpus callosum. Other features were growth retardation, hypertelorism, anomalies of the eyes, in part consistent with Peters' anterior chamber anomalies, mild brachymelia, brachydactyly, and also internal anomalies. Suspected PPS was confirmed by detection of B3GALTL mutation in these four fetuses and in one additional sib fetus, revealing homozygosity for the common c.660 + 1G > A donor splice site mutation in intron 8. CONCLUSIONS: Autosomal-recessive PPS has not yet been diagnosed prenatally. We want to alert ultrasonographers to the diagnosis of this disorder in growth-retarded fetuses with (recurrent) hydrocephaly, agenesis of the corpus callosum, and cleft lip/palate and stress the more severe fetal manifestation, describing a first such case with additional Dandy-Walker cyst and occult meningoencephalocele. © 2012 John Wiley & Sons, Ltd.

Approaches to Basal Ganglia Cavernomas

Infliximab As a Rescue-treatment in Difficult-to-treat Autoimmune Hepatitis

BACKGROUND: Autoimmune hepatitis is a chronic inflammatory liver disease that leads to liver cirrhosis and corresponding complications if left untreated. Current standard treatment with azathioprine and prednisolone induces remission in the vast majority of patients. However, for those patients not responding to standard treatment or not tolerating these drugs, few alternatives can be used and their effectiveness might be limited. OBJECTIVE: We sought to analyze the safety and efficacy of off-label treatment with infliximab in a cohort of eleven patients with difficult-to-treat autoimmune hepatitis. PATIENTS AND METHODS: Patients with difficult-to-treat autoimmune hepatitis who could not be brought into remission with standard treatment - either due to drug intolerance or due to insufficient drug impact - were treated off-label with infliximab for a minimum of six months. Patient files were reviewed retrospectively. RESULTS: Treatment with infliximab led to reduction in inflammation, evidenced by a decrease in transaminases (mean AST prior treatment 475 U/l +/- 466, mean AST during treatment 43 U/l +/- 32) as well as in immunoglobulins (pretreatment mean IgG 24,8 mg/dl +/- 10,1, mean IgG during treatment 17,38 mg/dl +/-6). Infectious complications occurred in seven out of eleven patients and close monitoring was necessary. CONCLUSION: Infliximab may be considered as rescue therapy in patients with difficult-to-treat autoimmune hepatitis albeit treatment may be associated with infectious complications.

Direct in Vivo Observations of P-selectin Glycoprotein Ligand-1-mediated Leukocyte-endothelial Cell Interactions in the Pulmonary Microvasculature in Abdominal Sepsis in Mice

OBJECTIVE: P-selectin glycoprotein ligand-1 (PSGL-1) has been shown to play a significant role in septic lung injury. However, the detailed role of PSGL-1 in the pulmonary leukocyte recruitment remains elusive. We have developed a method based on intravital fluorescence microscopy of the lung microcirculation to examine the role of PSGL-1 in the extravasation process of leukocytes in septic lung damage. METHODS: Male C57BL/6 mice were treated with a control antibody or an anti-PSGL-1 antibody prior to cecal ligation and puncture (CLP). Leukocyte-endothelium interactions and microvascular hemodynamics were studied in pulmonary arterioles, capillaries and venules 4 h after CLP. RESULTS: Immunoneutralization of PSGL-1 decreased CLP-induced leukocyte rolling in pulmonary arterioles and venules significantly. Inhibition of PSGL-1 had no effect on leukocyte adhesion in venules, whereas the number of adherent leukocytes in lung arterioles and the number of trapped leukocytes in capillaries were markedly decreased. Moreover, immunoneutralization of PSGL-1 improved microvascular perfusion in the lung of septic animals. CONCLUSIONS: Taken together, these results document that PSGL-1 mediates leukocyte rolling in arterioles and venules. However, inhibition of PSGL-1 only decreases leukocyte adhesion in arterioles, suggesting that leukocyte rolling is not a prerequisite for pulmonary venular adhesion of leukocytes in sepsis. In addition, our data show that capillary trapping of leukocytes is dependent on PSGL-1 function.

Chemical- and Effect-oriented Exposomics: Three Gorges Reservoir (TGR)

Exposomic studies of the rapidly changing environment of the Three Gorges Reservoir (TGR) after its impounding is elaborated as a novel field of human and environmental research. Molecular exposomics is focused on the measure of all exposures to molecules and especially persistent organic pollutants-like compounds are of emerging interest due to their lifetime existence in the environment and humans. Theoretical considerations in general and particular for the TGR are deduced and presented using quantitative approaches for this research field. Since exposomics is strongly time-dependent, a theory is presented to link extension of exposure, time, and related effects. Similarity to the first law of thermodynamics is outlined. On top of this, the integrated use of biomarkers is presented employing chemical analysis for biomarkers of exposure and effects, biomarkers in vivo, in vitro approaches and the link between chemical mixtures, and the onset of disease and lethality. Besides real organisms, also virtual organisms are favored to act as well-defined sub-compartments such as fat of biota and with respect to time of exposure. Exposomics is the perspective of risk evaluation and chronic exposures in the running century. It needs novel theories, approaches, and integrated action between medical and environmental disciplines. The existing knowledge about molecular stressors has to be assembled and put into a context especially with respect not only to time resp. lifetime exposure of humans but also eco-toxicological findings by using highly conserved phylogenetic mechanisms to enable links between human and risks of environmental biota. The TGR is a good example not only to employ biomonitoring of real but also virtual organisms due to the lack of established ecotopes in this changing environment so far. Progress in understanding long-term risks requires a proper theory as well as novel tools such as virtual organisms. On top, multidisciplinary approaches and the utilization of existing knowledge about the exposure of the environment and humans have to be merged and directed into mutual concepts. Effect-oriented and chemical analysis must be designed time-oriented to determine lifetime exposures of mankind and nature. Perspectively, a first attempt about exposomic theory and concepts is proposed and has to be developed experimentally further enclosing virtual besides of real organisms and compartments. Environmental and human exposomics have to be considered as a unified global issue in order to effectively utilize their mutual existing knowledge most effectively. The TGR is a challenging model system aiming this objective.

Support Vector Machine Classification Based on Correlation Prototypes Applied to Bone Age Assessment

Bone age assessment (BAA) on hand radiographs is a frequent and time consuming task in radiology. We present a method for (semi)automatic BAA which is done in several steps: (i) extract 14 epiphyseal regions from the radiographs, (ii) for each region, retain image features using the IRMA framework, (iii) use these features to build a classifier model (training phase), (iv) evaluate performance on cross validation schemes (testing phase), (v) classify unknown hand images (application phase). In this paper, we combine a support vector machine (SVM) with cross-correlation to a prototype image for each class. These prototypes are obtained choosing one random hand per class. A systematic evaluation is presented comparing nominal- and real-valued SVM with k nearest neighbor (kNN) classification on 1,097 hand radiographs of 30 diagnostic classes (0 19 years). Mean error in age prediction is 1.0 and 0.83 years for 5-NN and SVM, respectively. Accuracy of nominal- and real-valued SVM based on 6 prominent regions (prototypes) is 91.57% and 96.16%, respectively, for accepting about two years age range.

Evaluation of Noninvasive Follow-up Methods for the Detection of Intracranial In-Stent Restenosis: A Phantom Study

OBJECTIVES: Intra-arterial digital subtraction angiography (IA-DSA), an invasive procedure, is the current reference examination after percutaneous transluminal angioplasty and stenting for the detection of in-stent restenosis (ISR). In this phantom study, we evaluated flat-panel angiographic computed tomography after intravenous contrast agent application (IV-ACT) and multidetector computed tomographic angiography (MDCTA) as potential noninvasive follow-up alternatives after intracranial percutaneous transluminal angioplasty and stenting. MATERIALS AND METHODS: We simulated an intracranial vessel using a silicon tube placed inside a human skull. Three different stent systems were deployed inside the silicon tubes, each with diameters of 3 or 4 mm. Three grades of ISR (25%, 50%, and 75%) were simulated. The IA-DSA and IV-ACT examinations were performed on a flat-panel detector angiography system. The MDCTA images were acquired with a 128-slice computed tomographic scanner. The mean stenosis diameters, measured with each technique, were compared using the Bland-Altman plot. The difference between the known stenosis diameter and the measured stenosis diameter was calculated for each examination. RESULTS: Stenosis measurements on the IA-DSA images showed no statistically significant differences compared with the known stenosis diameters (P = 0.19). In the 3-mm stent category, when compared with the known stenosis diameter, mean (SD) differences of 0.01 (0.15) mm, 0.03 (0.24) mm and 0.16 (0.5) mm were calculated for the IA-DSA, IV-ACT, and MDCTA stenosis measurements, respectively. As for the 4-mm stents, IA-DSA and IV-ACT were again very accurate, with mean (SD) differences of -0.03 (0.11) mm and 0.07 (0.19) mm, respectively, compared with the known stenosis diameters, whereas MDCTA overestimated ISR, with a mean (SD) difference of 0.49 (0.53) mm. The Bland-Altman plots show a mean (SD) difference of 0.08 (0.2) mm between IA-DSA and IV-ACT (95% confidence interval, 0.05-0.11) and a mean (SD) difference of 0.34 (0.56) mm between IA-DSA and MDCTA measurements (95% confidence interval, 0.25-0.42). CONCLUSIONS: In our phantom study, IA-DSA was the only examination to predict accurately degrees of stenosis compared with the known stenosis diameters. The results of the IV-ACT measurements were comparable with those of IA-DSA. Multidetector computed tomographic angiography was less accurate in the quantification of stenosis, usually overestimating ISR.

Influence of PEEP on Cerebral Blood Flow and Cerebrovascular Autoregulation in Patients With Acute Respiratory Distress Syndrome

BACKGROUND:: High levels of positive end-expiratory pressure (PEEP), as part of the treatment in patients with acute respiratory distress syndrome (ARDS), may prevent alveolar collapse and maintain oxygenation. PEEP potentially reduces cerebral venous return, increases intracranial blood volume, and may, therefore, affect cerebral blood flow (CBF) and cerebrovascular autoregulation (AR). This study investigates the effect of PEEP on CBF and AR in patients with respiratory failure. METHODS:: CBF velocity was measured using transcranial doppler and correlated with the invasive arterial blood pressure curve to calculate the index of AR Mx (Mx>0.3 indicates impaired AR). Mx was measured at lower PEEP levels and after increasing PEEP. Only an increase of Mx of >0.2 was considered to be clinically relevant. Two 1-sided Wilcoxon tests. RESULTS:: Twenty mechanically ventilated patients with ARDS were included. Elevation of PEEP from 9.2±1 to 14.3±1 cm H2O did not influence CBF velocity but increased Mx from 0.317±0.35 to 0.414±0.32 (difference ≤0.2). Mx was >0.3 in 11/20 patients during baseline measurements, indicating impaired AR. CONCLUSIONS:: Surprisingly, AR was impaired in 55% of the patients with ARDS. This should be taken into account when managing cerebral perfusion pressure to avoid cerebral hyperperfusion or hypoperfusion. Increasing PEEP from 9.2 to 14.3 cm H2O had no further clinically relevant effect on AR, independent of preexisting AR impairment.

Increased ABA Sensitivity Results in Higher Seed Dormancy in Soft White Spring Wheat Cultivar 'Zak'

As a strategy to increase the seed dormancy of soft white wheat, mutants with increased sensitivity to the plant hormone abscisic acid (ABA) were identified in mutagenized grain of soft white spring wheat "Zak". Lack of seed dormancy is correlated with increased susceptibility to preharvest sprouting in wheat, especially those cultivars with white kernels. ABA induces seed dormancy during embryo maturation and inhibits the germination of mature grain. Three mutant lines called Zak ERA8, Zak ERA19A, and Zak ERA19B (Zak ENHANCED RESPONSE to ABA) were recovered based on failure to germinate on 5 μM ABA. All three mutants resulted in increased ABA sensitivity over a wide range of concentrations such that a phenotype can be detected at very low ABA concentrations. Wheat loses sensitivity to ABA inhibition of germination with extended periods of dry after-ripening. All three mutants recovered required more time to after-ripen sufficiently to germinate in the absence of ABA and to lose sensitivity to 5 μM ABA. However, an increase in ABA sensitivity could be detected after as long as 3 years of after-ripening using high ABA concentrations. The Zak ERA8 line showed the strongest phenotype and segregated as a single semi-dominant mutation. This mutation resulted in no obvious decrease in yield and is a good candidate gene for breeding preharvest sprouting tolerance.

Intrinsic Instability of Aberration-corrected Electron Microscopes

Aberration-corrected microscopes with subatomic resolution will impact broad areas of science and technology. However, the experimentally observed lifetime of the corrected state is just a few minutes. Here we show that the corrected state is intrinsically unstable; the higher its quality, the more unstable it is. Analyzing the contrast transfer function near optimum correction, we define an "instability budget" which allows a rational trade-off between resolution and stability. Unless control systems are developed to overcome these challenges, intrinsic instability poses a fundamental limit to the resolution practically achievable in the electron microscope.

Human Exposure to Fluorotelomer Alcohols, Perfluorooctane Sulfonate and Perfluorooctanoate Via House Dust in Bavaria, Germany

This study aimed at investigating the presence and distribution of fluorotelomer alcohols (FTOHs), perfluorooctane sulfonate (PFOS) and perfluorooctanoate (PFOA) in house dust to evaluate human exposure to these compounds via dust ingestion. 31 house dust samples were collected from Bavaria, Germany and analyzed for 4:2, 6:2, 8:2 and 10:2 FTOH, PFOS and PFOA. PFOA was the dominant compound in 79% of the dust samples, followed by PFOS and 8:2 FTOH, while 4:2 FTOH was not detected in any samples. The total concentration of per- and polyfluorinated compounds (PFCs) varied from 32.2 to 2456ng/g. In addition, the total ingestion rate for PFCs was 0.4-135ng/d for adults and 5.1-246ng/d for toddlers, and the highest 8:2 FTOH-based PFOA intake via indoor dust was 0.24ng/d for adults and 0.44ng/d for toddlers. Overall, the results of this study demonstrate that dust ingestion is a minor pathway for human exposure to these PFCs; the PFC ingestion via indoor dust is generally low, and only under a worst scenario high intakes have to be expected for toddlers.

Next-generation RNA Sequencing Reveals Differential Expression of MYCN Target Genes and Suggests the MTOR Pathway As a Promising Therapy Target in MYCN-amplified Neuroblastoma

In many cancer types, MYC proteins are known to be master regulators of the RNA-producing machinery. Neuroblastoma is a tumor of early childhood characterized by heterogeneous clinical courses. Amplification of the MYCN oncogene is a marker of poor patient outcome in this disease. Here, we investigated the MYCN-driven transcriptome of 20 primary neuroblastomas with and without MYCN amplification using next-generation RNA sequencing and compared the results to those from an in vitro cell model for inducible MYCN (SH-EP MYCN-ER). Transcriptome sequencing produced 30-90 million mappable reads for each dataset. The most abundant RNA species was mRNA, but snoRNAs, pseudogenes and processed transcripts were also recovered. A total of 223 genes were significantly differentially expressed between MYCN-amplified and single-copy tumors. Of those genes associated with MYCN both in vitro and in vivo, 32% of MYCN upregulated and 37% of MYCN downregulated genes were verified either as previously identified MYCN targets or as having MYCN-binding motifs. Pathway analyses suggested transcriptomal upregulation of mTOR-related genes by MYCN. MYCN-driven neuroblastomas in mice displayed activation of the mTOR pathway on the protein level and activation of MYCN in SH-EP MYCN-ER cells resulted in high sensitivity toward mTOR inhibition in vitro. We conclude that next-generation RNA sequencing allows for the identification of MYCN regulated transcripts in neuroblastoma. As our results suggest MYCN involvement in mTOR pathway activation on the transcriptional level, mTOR inhibitors should be further evaluated for the treatment of MYCN-amplified neuroblastoma.

Genistein and Dicarboximide Fungicides in Infant Formulae from the EU Market

A method based on ultrasonic extraction and purification by solid phase extraction followed by LC-MS/MS and GC-MS analysis was developed for the determination of genistein, genistin, iprodione, vinclozolin and procymidone in infant powdered formulas. The method was tested for different formulations: milk, soy and hypoallergenic, and was applied to European pooled samples. Spike recoveries ranged from 53.1% to 91.5% and the relative standard deviation values for repeatability ranged from 9.6% to 17.7%, except for iprodione in milk formula (22.3%). None of the fungicides were found in the European pooled formulae, while genistein was found at 9.7μg/g in soy formula and the concentration of genistin, its β-glycosylated form, was respectively 31.4ng/g and 476ng/g in milk and soy formula.

PET/MRI in Head and Neck Cancer: Initial Experience

To evaluate the feasibility of PET/MRI (positron emission tomography/magnetic resonance imaging) with FDG ((18)F-fluorodeoxyglucose) for initial staging of head and neck cancer.

Waiting
simple hit counter