Emergency catheter ablation is justified in critical patients with drug-refractory life-threatening arrhythmias. The procedure can be used for ablation of an accessory pathway in preexcitation syndrome with high risk of ventricular fibrillation and in patients with shock due to ischemic cardiomyopathy and incessant ventricular tachycardia. Emergency catheter ablation can also be justified in patients with an electrical storm of the implanted cardioverter-defibrillator or in patients with idiopathic ventricular fibrillation. Performance of catheter ablation as an emergency procedure in critical patients is rare. Catheter ablation is indicated in patients with drug-refractory life-threatening arrhythmias. The scenarios discussed below will illustrate the importance of emergency catheter ablation in preventing fatality.Sudden cardiac death can occur in patients with Wolff-Parkinson-White (WPW) syndrome due to rapid antegrade conduction of atrial fibrillation over an accessory pathway, with resulting ventricular fibrillation and cardiac arrest.et al. reported on 48 patients with WPW syndrome and aborted sudden cardiac death. In all patients, the accessory pathways were successfully ablated and ICD implantation was not performed. During the follow-up period of 5 ± 2 years there was no recurrence of life-threatening arrhythmia or syncope. Therefore catheter ablation of the accessory pathway is the appropriate treatment in this group of patients.A 31-year-old female patient was resuscitated by her husband after the latter recognized syncope and cessation of breathing. The emergency physician diagnosed ventricular fibrillation and immediate defibrillation resulted in restoration of normal sinus rhythm. The patient was intubated, ventilated, and transferred to the nearest hospital. After 3 days of intensive care therapy, including mild hypothermia, extubation was successfully performed. She had no neurological deficits. A 12-lead surface electrocardiogram (ECG) showed preexcitation syndrome with a suspected accessory pathway at the inferoseptal aspect. The patient was then referred to our hospital for catheter ablation. During mapping of the inferoseptal region of the right atrium close to the coronary sinus ostium, the accessory pathway could be located slightly inside the coronary sinus. Therefore mapping was performed via transaortic access and the accessory pathway was successfully ablated at the atrial aspect of the mitral annulus . An implPatients with ischemic or nonischemic cardiomyopathy can suffer from so-called incessant ventricular tachycardias (VT). These tachycardias are often slow and therefore incessant and drug refractory. These arrhythmias occur in the setting of severe depressed left ventricular function with large areas of scar tissue. The patients are often on antiarrhythmic drugs  and sometimes present in cardiogenic shock and have to be treated in intensive care units. The treatment is termination of VT by electrical cardioversion. Additional antiarrhythmic drugs cannot be tolerated as they have negative inotropic action and worsen the patient's condition. In this setting, catheter ablation is the only option for these patients.® mapping, the reentry circuit could be identified at the free wall of the right ventricle. Entrainment maneuvers in areas with mid-diastolic potentials were found positive and multiple radiofrequency current applications could terminate the VT. Ablation was started with an 8-mm solid-tip electrode. Later in the procedure, a 4-mm cooled-tip electrode was used. After 36 h in intensive care the VT recovered and a second ablation procedure was performed, with successful termination of VT and restoration of sinus rhythm. The patient recovered quickly, blood pressure returned to normal, and edema and elevated liver enzymes normalized within the next 3 days.A 63-year-old male patient with arrhythmogenic right ventricular cardiomyopathy was referred from another hospital for catheter ablation of incessant VT. Antiarrhythmic drug treatment consisted of a beta-blocker and amiodarone. An ICD had been implanted some years ago after aborted sudden cardiac death. During electrophysiologic study, VT was confirmed . After rThe so-called electrical storm with multiple ICD discharges due to VT or VF is life threatening, and sudden cardiac death can occur despite the ICD. Amiodarone and β-blockers are almost always given as first-line treatment to terminate VT or VF. In a recent study by Carbucicchio in 95 patients , catheter ablation was performed in drug-refractory electrical storm.[The success rate of catheter ablation for acute treatment of electrical storm in our study is high; similar high rates have also been reported by Schreieck and others. One example of our experience is given in Figures –6. CatheA rather small number of patients in our study had so-called idiopathic VF. Most of them had aborted sudden cardiac death and later had had an ICD implanted. Haissaguerre reported on 27 patients with idiopathic VF without apparent structural heart disease. The uniqFrom our cases and the results of our interventions we feel that emergency catheter ablation is justified in the following conditions: 1) Wolff-Parkinson-White (WPW) syndrome with risk of sudden cardiac death, 2) incessant VT due to ischemic cardiomyopathic shock, 3) electrical storm in patients with ICDs, and 4) idiopathic VF.


Acoels are simply organized unsegmented worms, lacking hindgut and anus. Several publications over recent years challenge the long-held view that acoels are early offshoots of the flatworms. Instead a basal position as sister group to all other bilaterian animals was suggested, mainly based on molecular evidence. This led to the view that features of acoels might reflect those of the last common ancestor of Bilateria, and resulted in several evo-devo studies trying to interpret bilaterian evolution using acoels as a proxy model for the "Urbilateria".Symsagittifera roscoffensis. Gene content and circular organization of the mitochondrial genome does not significantly differ from other bilaterian animals. However, gene order shows no similarity to any other mitochondrial genome within the Metazoa. Phylogenetic analyses of concatenated alignments of amino acid sequences from protein coding genes support a position of Acoela and Nemertodermatida as the sister group to all other Bilateria. Our data provided no support for a sister group relationship between Xenoturbellida and Acoela or Acoelomorpha. The phylogenetic position of Xenoturbella bocki as sister group to or part of the deuterostomes was also unstable.We describe the first complete mitochondrial genome sequence of a member of the Acoela, Our phylogenetic analysis supports the view that acoels and nemertodermatids are the earliest divergent extant lineage of Bilateria. As such they remain a valid source for seeking primitive characters present in the last common ancestor of Bilateria. Gene order of mitochondrial genomes seems to be very variable among Acoela and Nemertodermatida and the groundplan for the metazoan mitochondrial genome remains elusive. More data are needed to interpret mitochondrial genome evolution at the base of Bilateria. Acoels are marine, soft-bodied, unsegmented worms without hindgut and anus - the mouth opens to a central digestive parenchyma, a gut lumen is absent. Acoels move with their multiciliated epidermis although many are 'surprisingly muscular' . Most ofEarly molecular systematic studies using ribosomal RNA genes strongly suggested that Acoela and Nemertodermatida were distinctly separate from the Platyhelminthes -14; thisIsodiametra pulchra . CREx . CREx  was usedatp8 and nad4l, as these are the shortest and least conserved genes from the protein-coding set of animal mt genomes. A total of 60 species was chosen to build the alignments. A detailed overview of the respective taxa and their accession numbers is given in table http://www.cgae.de. NH-PhyloBayes [For the phylogenetic analyses we concatenated amino acid alignments from 11 protein-coding genes. We omitted alignments from yloBayes  was usedatp 6/8: ATPase subunit 6/8 genes; cob: cytochrome b gene; cox 1-3:cytochrome oxidase subunit I-III genes; mtDNA: mitochondrial DNA; nad1-6 and nad4L: NADH dehydrogenase subunit 1-6 and 4L genes; rrnS/rrnL: small/large rRNA subunit genes; rRNA: ribosomal RNA; tRNA: transfer RNA; trnX: tRNA gene X ('X' replaces the one-letter amino acid code of the respective tRNA); nt: nucleotides; PCR: polymerase chain reactionAM did most of the laboratory experiments. XB reared animals and provided helpful EST data. All authors analysed parts of the data and did phylogenetic analyses. All authors discussed results. AM, LP and DTJL wrote substantial parts of the manuscript. All authors read and approved the final manuscript.Supplemental Figs. S1-3. Gene order comparison with CREX ; phylogenetic analysis under exclusion of Acoela and Nemertodermatida; phylogenetic analysis under exclusion of Xenoturbella.Click here for file


The cell-surface proteins of 6 different melanoma cell cultures have been labelled with 125I using lactaperoxidase-catalysed iodination. Fractionation of the proteins was achieved using 5--22.5% polacrylamide-gradient gel electrophoresis in the presence of sodium dodecyl sulphate (SDS) and the proteins were detected by autoradiography. Up to 24 labelled proteins were detected in the individual cell cultures, but the proteins labelled differed considerably in the 6 cultures examined. A possible reason for this, involving variation in the glycosylation of cell-surface glycoproteins is discussed. Cells of the same melanoma line had similar cell-surface proteins at different passage levels, but changes in the labelled proteins occurred when the culture conditions were altered. The cell-surface proteins of high molecular weight were cleaved by trypsin, but most of the low mol.-wt. proteins were resistant to trypsin. The "large external transformation sensitive" (LETS) protein detected as a major protein on fibroblasts in culture was not a dominant protein on the melanoma cells. It was detected on only 4/6 cell cultures. Possible relationships of the cell-surface proteins described in this study to morphology, immunological properties and proteolytic activity of human melanoma cells are discussed.


These chitosan based polyelectrolyte complexes exhibit favourable physicochemical properties with preservation of chitosan’s biocompatible characteristics. These complexes are therefore good candidate excipient materials for the design of different types of dosage forms. It is the aim of this review to describe complexation of chitosan with selected natural and synthetic polyanions and to indicate some of the factors that influence the formation and stability of these polyelectrolyte complexes. Furthermore, recent investigations into the use of these complexes as excipients in drug delivery systems such as nano- and microparticles, beads, fibers, sponges and matrix type tablets are briefly described.Chitosan has been the subject of interest for its use as a polymeric drug carrier material in dosage form design due to its appealing properties such as biocompatibility, biodegradability, low toxicity and relatively low production cost from abundant natural sources. However, one drawback of using this natural polysaccharide in modified release dosage forms for oral administration is its fast dissolution rate in the stomach. Since chitosan is positively charged at low pH values (below its pK Mixing oppositely charged polyelectrolytes in solution will result in their self assembly or spontaneous association due to the formation of strong, but reversible electrostatic links. These direct interactions between the polymeric chains lead to the formation of polyelectrolyte complex networks with non-permanent structures while avoiding the use of covalent cross-linkers. In general, these polymeric networks or hydrogels are well tolerated, biocompatible and are more sensitive to changes in environmental conditions ,2. The fN-acetyl-d-glusoamine (acetylated units)  and d units)  with typThe cationic amino groups on the C2 position of the repeating glucopyranose units of chitosan can interact electrostatically with the anionic groups  of other polyions to form polyelectrolyte complexes. Many different polyanions from natural origin  or synthetic origin ), polyphosphoric acid, poly (L-lactide) have been used to form polyelectrolyte complexes with chitosan in order to provide the required physicochemical properties for the design of specific drug delivery systems .,29.23,29It was shown that the nature or type of carrageenan considerably influence the characteristics of the polyelectrolyte complex that is formed with chitosan. The mechanical strength of polyelectrolyte complex gels formed between chitosan and different carrageenans were in the order λ- > ι- > κ-carrageenan and although the latter two formed stronger gels due to the formation of more cross-links as a result of their double helix secondary structures, these gels were also more brittle. Furthermore, the gels obtained for ι- and κ-carrageenan were temperature sensitive because of the helix coil conformational transitions in their molecules .An investigation was done on the potential of polyelectrolyte complexes between κ-, ι-, λ-carrageenan and chitosan to form controlled release systems for glucose oxidase. The complex between chitosan and κ-carrageenan showed high encapsulation efficiencies for glucose oxidase while having the lowest release rate for this compound. Furthermore, this complex was able to protect the encapsulated glucose oxidase against degradation in pH 1.2 solution, in a chitosanase solution and in a pepsin solution .The polysaccharides of the plant cell wall consist mainly of cellulose, hemicelluloses and pectin. Pectin is a linear polysaccharide composed of α-1,4-linked D-galacturonic acid units, however, this linear structure is interrupted with highly branched regions in the polymer chain . The comWhen acidic chitosan and pectin solutions are mixed a homogenous solution is obtained without any ionic interactions between the two polymers. A polyelectrolyte complex could be obtained by adjusting the pH of this mixture to a value of 5.5 where electrostatic interactions between the negatively charged carboxylic acid groups of pectin and the positively charged amino groups of chitosan occurred. As expected, the extent of this interaction depended on the pH of the surrounding medium which determined the extent of ionization of the polymers .Polyelectrolyte complexes composed of chitosan and pectin were compressed into tablets with vancomycin as model drug. The tablets showed a pH sensitive swelling ability with drug release behaviour based on enzyme dependent degradation in the presence of beta-glucosidase, which indicated potential for colon-specific delivery of vancomycin .Xanthomonas campestris. It consists of a cellulosic backbone, namely β--d-glucopyranose glucan, with a trisaccharide side chain, namely -α-d-mannopyranose--β-d-glucuronic acid--β-d-mannopyranose, on every second glucose residue -d-glucuronic acid repeating units linked by β (1→3) bonds  is the only nonsulfated glycosaminoglycan found in the extracellular matrix throughout connective, epithelial and neural tissues. Hyaluronic acid is a linear anionic polysaccharide with high molecular weight that consists of β -3) bonds . It is pIt was shown that the polyelectrolyte complex between chitosan and hyaluronic acid protected hyaluronic acid against enzymatic hydrolysis, but only at pH values different from the optimal pH of the enzyme. The results from this study revealed that the chitosan-hyaluronic acid polyelectrolyte complex unfortunately had less cell proliferation and wound healing effects compared to chitosan alone .N-isopropylacrylamide) it prevented disintegration of the system during in vitro drug release studies and thereby facilitated controlled release of nalbuphine [Chitosan-hyaluronic acid polyelectrolyte complex beads were prepared and these beads were reacted in another step with hyaluronic acid to form an outer layer around the bead, which increased their stability . Furtherlbuphine .Chitosan-hyaluronic acid polyelectrolyte complex nanoparticles loaded with heparin were effectively internalised by rat mast cells and showed similar activity in terms of preventing histamine release compared to free heparin. A high degree of loading efficiency was observed (approximately 70%) and the nanoparticles were stable in phosphate buffered saline for at least 24 h. The release of unfractionated heparin was much slower (10.8% within 12 h) than that of low molecular weight heparin (79.7% within 12 h). These results indicated the usefulness of these nanoparticles to serve as effective delivery systems for pulmonary administration .Cochlospermum gossypium, a tree growing naturally in India. It consists of a complex polysaccharide with a relatively high molecular weight (ranging between 9.8 × 105 and 2.5 × 107 g/mol) and therefore exhibit a high viscosity in solution with gelation characteristics. When the acid hydrolysed gum was characterised by gas chromatography linked to mass spectrometry (GC-MS), the following monosaccharides were indicated to be present: rhamnose, galacturonic acid, glucuronic acid, β-d-galactopyranose, α-d-glucose, galactose, arabinose, mannose and fructose [Gum kondagogu is a natural biopolymer that is obtained from the exudated gum of fructose ,48.max) was higher compared to that of free dicolfenac sodium, the extent of drug absorption (area under the concentration-time curve) was also increased significantly when administered in the form of the microparticles [Chitosan-gum kondagogu polyelectrolyte complex microparticles loaded with diclofenac sodium showed controlled drug release, which was depended on the pH of the dissolution medium. A pharmacokinetic study of these microparticles in rats revealed that although the time to reach the peak plasma concentration  and below 6.2 (above which chitosan start to precipitate out of solution) . ChitosaGamma poly (glutamic acid) is a hydrophilic polyanion that is water-soluble, non-toxic and biodegradable. This polyamino acid is formed by amide bond linkages between the amino group on theα carbon atom of one glutamic acid molecule and the carboxyl group on the γ carbon atom of the next glutamic acid molecule .It was shown that stable nano-sized particles can be formed as a direct result of interactions between the carboxylic groups of linear γ-poly (glutamic acid) chains and the amino groups of linear chitosan. The size and stability of the nanoparticles depended on factors such as medium pH, concentration and ratio of γ-poly (glutamic acid) and chitosan solutions as well as the mixing order .in vivo studies that investigated delivery of encapsulated insulin, the nanoparticles were formed by self-assembly of chitosan and γ-poly (glutamic acid) in the presence of the cross-linkers such as tripolyphosphate and magnesium sulphate.In a study wherein chitosan-γ-poly (glutamic acid) polyelectrolyte complex nanoparticles were investigated as drug carriers for targeted drug delivery, it was shown that they can penetrate A2780/AD ovarian cancer cells and this penetration was significantly faster and more efficient in the presence of conjugated folic acid due to over expression of folate receptors on these cells . It was Maleic starch half-ester acid is an anionic derivative of starch that produces a polyelectrolyte complex when combined with the polycation, chitosan. Characterisation and evaluation of this complex indicated the driving force for self-assembly was electrostatic interactions. A pH sensitive release of 5-fluorouracil was obtained that was also dependent on the maleic starch half-ester to chitosan ratio as well as the carboxyl content of maleic starch half-ester used as starting material .®) and Carbopol are high molecular weight polymers consisting of acrylic acid monomers. The poly (acrylic acid) chains are cross-linked with divinyl glycol to form polycarbophil and polyalkenyl alcohols such as allyl ethers of pentaerythritol or allyl ethers of sucrose to form Carbopol. Cross-linking of the poly (acrylic acid) chains with divinyl glycol in the case of polycarbophil renders only 20% of the carboxylic acid groups inaccessible for interactions with other compounds [Polycarbophil  as well as between chitosan and Carbopol 71G NF®, which showed promising characteristics in terms of flexibility, resistance and water vapour transmission rate. These films showed such great potential as formulations for topical and transdermal drug administration during this characterization study that it was suggested they should be investigated for incorporation, release and delivery of drugs [Films for dermal drug delivery were prepared from the polyelectrolyte complexes between chitosan and polycarbophil (Noveon AA-1of drugs .®) polyelectrolyte complex as matrix former approached zero order release for two model drugs with different water solubilities . The rate of the release of these two model drugs depended on the concentration of the polyelectrolyte complex in the formulation of the matrix systems. When the matrix type tablets consisted only of polyelectrolyte complex (without drug), they showed a very high swelling capability without erosion, which means the swelling is completely reversible upon drying without any weight loss [®) were capable of slower drug release than those prepared from hydroxypropylmethylcellulose [®) polyelectrolyte complex, it was shown that the mechanism of release was dependent on the concentration of the polyelectrolyte complex in the formulation of the matrix systems relative to the other excipients. The matrix systems which contained the highest concentration of the polyelectrolyte complex approached zero-order insulin release and exhibited erosion in conjunction with swelling [in vitro drug release results.Monolithic matrix type tablets prepared from chitosan-polycarbophil (Noveon AA-1ght loss . In anotellulose . When inswelling . In genePolymethacrylate copolymers (or Eudragit) are well known film-coating agents for oral dosage forms such as capsules or tablets. Different types of Eudragit polymers exist which are synthetic methacrylic acid copolymers consisting of different ratios of dimethylaminoethylmethacrylates, methacrylic acid and methacrylic acid esters. Some of them are polycations  due to the presence of dimethylamino groups or quaternary amino groups, while others are polyanions (Eudragit L and S) due to the presence of carboxylate groups ,66.The first reference to an ionic interaction between chitosan and Eudragit was in a study where chitosan microcores containing sodium diclofenac as model drug were microencapsulated in Eudragit S-100 using an oil-in-oil solvent evaporation technique for colon targeted release. The ionic interaction between the two polymers was confirmed by means of infrared spectroscopy and had an influence on the drug release profile from the microspheres .Polyelectrolyte complexes formed between chitosan with different molecular weights and Eudragit L100 or Eudragit L100-55 were compressed into matrix type tablets containing diclofenac sodium as model drug. Different aspects of the complex systems were investigated such as the molar ratio, structure, swelling and drug release profiles. Results indicated that this polyelectrolyte complex has high potential for manufacture of controlled release drug delivery systems. Factors such as the composition of the polyelectrolyte complex in terms of polymer ratios, Eudragit type and molecular weight of the chitosan influenced the drug release rate from the matrix type tablets .By changing the alkyleneoxide chain composition, a series of polyelkylenoxide-maleic acid (PAOMA) copolymers can be synthesised with different characteristics  where i and j represent the random copolymerisation degree.The ethylene oxide (EO) ratio determines whether the copolymer is hydrophilic or hydrophobic. EO ratio means the molar fraction of ethylene oxide in alkylene oxide chain defined by The scope of polymers used in dosage form design can be increased by several approaches such as modification of their chemical structure, by combining different polymers in physical mixtures or by formation of polymer-polymer associations such as polyelectrolyte complexes. Polyelectrolyte complexes combine unique physicochemical properties of different polymers with the advantage of retaining high biocompatibility. It is therefore not surprising that polyelectrolyte complexes are gaining importance in modern pharmaceutical technology .in vitro studies conducted on chitosan-based polyelectrolyte complexes it is clear that they are valuable excipients with specific properties for efficient dosage form design, which may be valuable in the development of modified drug delivery systems. Unfortunately, the literature lacks in vivo data in terms of drug delivery from these dosage forms which makes it difficult to be conclusive in terms of their effectiveness as drug carriers at this stage. Since some work has been done on in vitro-in vivo correlations with chemically cross-linked chitosan hydrogels with successful sustained drug delivery in animals , it is


We present a 3D multi-cell simulation of a generic simplification of vascular tumor growth which can be easily extended and adapted to describe more specific vascular tumor types and host tissues. Initially, tumor cells proliferate as they take up the oxygen which the pre-existing vasculature supplies. The tumor grows exponentially. When the oxygen level drops below a threshold, the tumor cells become hypoxic and start secreting pro-angiogenic factors. At this stage, the tumor reaches a maximum diameter characteristic of an avascular tumor spheroid. The endothelial cells in the pre-existing vasculature respond to the pro-angiogenic factors both by chemotaxing towards higher concentrations of pro-angiogenic factors and by forming new blood vessels via angiogenesis. The tumor-induced vasculature increases the growth rate of the resulting vascularized solid tumor compared to an avascular tumor, allowing the tumor to grow beyond the spheroid in these linear-growth phases. First, in the linear-spherical phase of growth, the tumor remains spherical while its volume increases. Second, in the linear-cylindrical phase of growth the tumor elongates into a cylinder. Finally, in the linear-sheet phase of growth, tumor growth accelerates as the tumor changes from cylindrical to paddle-shaped. Substantial periods during which the tumor grows slowly or not at all separate the exponential from the linear-spherical and the linear-spherical from the linear-cylindrical growth phases. In contrast to other simulations in which avascular tumors remain spherical, our simulated avascular tumors form cylinders following the blood vessels, leading to a different distribution of hypoxic cells within the tumor. Our simulations cover time periods which are long enough to produce a range of biologically reasonable complex morphologies, allowing us to study how tumor-induced angiogenesis affects the growth rate, size and morphology of simulated tumors. Nutrient and waste diffusion lengths limit the size of such e.g. during wound healing and expansion of tissues during the female reproductive cycle.Both fetal and adult angiogenesis is primarily a response to hypoxia α DNA-binding activity in the nucleus varies exponentially with oxygen tension over the physiological range in mammalian cells α in their nuclei in less than 2 minutes α changes the expression levels of numerous hypoxia-dependent genes including those responsible for oxygen transport, vascular regulation, angiogenesis, glucose uptake, and glycolysis  and increases the stability of those boundaries contact inhibition of growth)Cell-adhesion plays a crucial role in the formation and stabilization of nascent blood vessels  continuum models include scales down to the grid size used to solve the equations. Continuum multi-grid and adaptive-mesh techniques can cover scales from Because of the range of scales involved in cancer biology, cancer simulations employ a wide range of techniques depending on their biological focus. Standard partial-differential-equation  and spread to other organs (metastasis) via blood vessels. The simple model we present in this initial paper neglects the crucial effects of detailed transport due to blood flow and the effects of flow-induced vascular remodeling. We discuss some of the resulting artifacts and missing behaviors below.The effects of blood flow on vascular remodeling and tumor growth have been extensively studied by Owen et al. GGH, also known as the Cellular Potts Model, CPM) in vitro. Rubenstein and Kaufman showed that invasion is maximized at intermediate collagen concentrations, as occurs experimentally.Because the Glazier-Graner-Hogeweg model (ECM) and smaller than those due to tissue structure. These inhomogeneities may affect tumor growth rates and morphology and the somatic evolution of metastatic potential within tumors. Thus, simulating vascular structure at the cell level is crucial to developing biomedically-meaningful tumor simulations.In our simulations, the vascular structure produces inhomogeneities in oxygen tension on scales larger than those appearing in continuum simulations depending on inhomogeneities in Extracellular Matrix , which behave identically to vascular cells until the concentration of VEGF-A exceeds an activation threshold. 10) We adopt the vascular-patterning hypothesis that activated vascular endothelial cells secrete and chemotax towards a short-diffusion-length chemoattractant , and lactic acid accumulation.In this paper we simulate 3D solid tumor growth and angiogenesis using the multi-cell GGH model. Our simulation omits many biological details, but provides a useful starting point for the construction of more realistic models. We focus on tumors where the vasculature remains peripheral to the growing tumor. Our major simplifications include: 1) We neglect the distinction between veins and arteries, anastomosis, and the possible presence of pericytes and smooth muscle cells. 2) Since oxygen-depleted areas of a tissue coincide with nutrient-depleted and energy-depleted areas Hypoxic domains form on day 8  would produce larger solid tumors both with and without angiogenesis. Since a tumor can grow to a maximum diameter of 200 µm without angiogenesis, average vascular branch lengths shorter than 200 µm will produce infinitely long cylindrical solid tumors.normal and hypoxic cells within the tumor also affects growth, reducing the inhomogeneities in cell proliferation which change spherical tumors into cylinders. Higher cell motilities paradoxically arrest tumor growth at diameters of 200 µm (no angiogenesis) and 300 µm (with angiogenesis) (data not shown). In contrast, lower cell motilities enhance formation of fingers and nodules which increase the invasiveness of the solid tumor. In reality, motility also depends on the adhesiveness of cells to each other and to the ECM, so this effect may not be clinically relevant.The random motility of We also assume that the vasculature remains peripheral to the tumor. Formation of blood vessels inside the tumor enhances oxygen transport and allows arbitrarily large tumors and faster, though not necessarily more invasive, growth.ECM in our simulation is not confined by physical boundaries and does not have a volume constraint, the pressure in our tumor does not change due to tumor growth. Explicitly modeling the cells and ECM in the peri-tumor region would result in a more realistic pressure.Tumor growth in real tissues leads to increasing hydrostatic and solid pressures, inducing tumor-cell quiescence and necrosis and also causing blood vessels to collapse. Because Our simplified tumor induces angiogenesis through VEGF signaling, but neglects other tumor-induced changes in the surrounding vasculature, including apoptosis of endothelial cells because of reduced pericyte coverage and lowered pH. Combining these vascular remodeling mechanisms with blood-flow calculations would give further insight into the effects of angiogenesis on vascular tumor growth.Although our high-resolution simulation represents a fairly small tissue volume, our results can scale to treat larger tissue volumes at lower spatial resolution. Such scaling tradeoffs may be useful for simulating tumors like prostate cancer, which involve multiyear progressions and centimeter-scale tissues.MPI) parallel implementation of the GGH which permits simulations with more than A real solid tumor with a volume of about 1 ll3D see  can simuOur next extension of these simulations will include blood flow to improve our description of substrate transport and also include shear effects on vascular remodeling.GGH, also known as the Cellular Potts Model, CPM), a multi-cell, lattice-based, stochastic model which describes biological cells and their interactions in terms of Effective Energies and constraints. GGH applications include modeling avascular tumor growth cell lattice of lattice sites, or voxels, at locations cell index, cell type, Effective Energy governs changes in the cell configuration. The basic Effective Energy describes constant-volume cells interacting via differential adhesion:i.e., deviation of Effective Energy), and modified Metropolis dynamics. For each attempt of a cell to displace a neighbor, we select at random a cell boundary (source voxel) and a neighboring target voxel and calculate the change in the effective energy i.e., the change is energetically favorable, we make it. If Monte Carlo Step (MCS). We describe secreted morphogens and oxygen macroscopically by concentration fields discretized at the resolution of the cell lattice. Fields evolve according to simple time-dependent partial-differential equations (PDEs). In addition to contact and volume energies, we include a chemotaxis term in the Effective Energy to model angiogenesis (see below).Our simulation uses the Glazier-Graner-Hogeweg model ((ECM). The total volume and surface area of ECM are not constrained. ECM voxels can be both source voxels, e.g. during retraction of the trailing-edge of a cell, and target voxels, e.g. during formation of lamellipodia. Since ECM does not have cytoskeletal fluctuations, we use the amplitude of cytoskeletal fluctuations of the target or source cell to determine the acceptance of the displacement.We define a special, generalized cell representing the extracellular medium normal,hypoxic and necrotic . Normal tumor cells become hypoxic when the oxygen partial pressure  and proliferate at a rate which depends on the oxygen partial pressure according to a Michaelis-Menten form. To model tumor cell proliferation, we increase the cell's target volume according to:maximum growth rate, doubling volume, we split the cell along the x–y plane. The two daughter cells receive equal target volumes of half of the target volume of the mother cell at mitosis. Necrotic cells lose volume at a constant rate:Our model contains three tumor-cell types: Hypoxic cells secrete the fieldHypoxic cells stop secreting VEGF-A and become normal if necrotic if Hypoxic cells secrete hypoxic cells and We remove zero-volume cells. endothelial cell (EC) types: vascular and neovascular. We further distinguish two types of neovascular cells, inactive neovascular and active neovascular.Vascular cells build the preexisting mature vasculature. To represent tight junctions between endothelial cells in mature capillaries which maintain the integrity of blood vessels, linear springs connect the centers-of-mass of vascular cells. The springs obey the elastic constraint, Inactive neovascular cells behave exactly like vascular cells. However, above a threshold value inactive neovascular cells switch irreversibly into active neovascular cells. Active neovascular cells can proliferate and chemotax up gradients of VEGF-A. To account for the contact-inhibited growth of neovascular cells, when the common surface area with other vascular, inactive neovascular and active neovascular cells is less than a threshold, we increase the target volume of the active neovascular cells according to the Michaelis-Menten relation:Active neovascular cells divide along the x–y plane when their volume reaches a specified doubling volume. After mitosis, both daughter cells are active neovascular and inherit half of their mother's target volume. Active neovascular cells at the tip of a sprout share two features with real tip-cells: 1) Compared to stalk cells, they have more free boundary which can respond to gradients of VEGF-A, dragging the rest of the sprout up the gradients. 2) Dragging reduces the contact area between active neovascular cells, promoting neovascular growth. Unlike real tip-cells, active neovascular cells at the tip of a sprout proliferate. Thus sprouts grow faster in steeper gradients of VEGF-A as long as the concentration of VEGF-A is well below the saturation concentration. We add a saturated Savil-Hogeweg-type chemotaxis term with contact inhibition to the basic field near the active neovascular cell's boundary s is a positive number which scales the VEGF-A concentration field relative to the neovascular activation threshold. Our model also contains two basic active neovascular cells up gradients of VEGF-A produce a dispersed growing population of neovascular cells rather than a connected capillary network vascular and neovascular cells into a capillary-like structure we extended to 3D Merks' 2D model of angiogenesis in which endothelial cells self-organize into capillary-like networks in response to autocrine chemotaxis to a very short-diffusing chemoattractant field by Vascular and neovascular cells secrete it at a constant rate ECM, and diffuses at a constant rate vascular and neovascular cells and vascular and neovascular cells in our model chemotax up gradients of Growth and chemotaxis of vascular cells makes the preexisting vasculature more stable and less flexible.Autocrine chemotaxis produces a capillary-like structure, while the elastic constraint between fieldneovascular cells form in closed loops, we assume that the oxygen partial pressure in the preexisting capillaries, field concentration neighboring a vessel changes proportionally to tumor cells. Since oxygen consumption of human cells is almost constant until Tumor cells take up oxygen at a rate proportional to ECM. We include oxygen consumption by EC cells in the adjusted oxygen partial pressure. Thus oxygen evolves according to:vascular and neovascular cells and normal and hypoxic cells. We have summarized the properties of the fields in Vasculature transports oxygen to the host tissue and the tumor. We represent the oxygen partial pressure, http://www.compucell3d.org/) tumor cell has a volume of about tumor cells in the simulation with angiogenesis on day 60 . We assume that both hypoxic and normal tumor cells in our model consume oxygen at a rate up to 3 times that of the cells in the surrounding tissue, thus tumor cells at We start our simulation with a single sels see . We assuneovascular cell that does not contact any vascular or neovacular cells has a minimum cell-cycle time of 24 h. Due to contact-inhibition of growth, neovascular cells incorporated in a tumor-induced vessel grow at a negligible rate.Experimentally, the VEGF-A diffusion constant is about 10 µmtumor cells and that cell-cell adhesion via cadherins keeps the tumor solid, i.e. that the surface tension at the tumor-ECM interface is positive, EC(vascular and neovascular) and tumor cells Click here for additional data file.Movie S1Initially, the tumor grows exponentially and reaches a maximum diameter of about 200 micrometers. The growing cylindrical tumor pushes and stretches the vessels it contacts, finally rupturing them on day 31. Total movie time = 75 days. Time interval between frames = 6 hours. The camera rotates at a rate of 0.1 degree per frame. Cell types: Green: normal; yellow: hypoxic; blue: necrotic; red: vascular. Axes are labeled in micrometer.(9.96 MB AVI)Click here for additional data file.Movie S2Initially, the tumor grows exponentially. When oxygen level drops below a threshold, the normal tumor cells become hypoxic and start secreting a long-diffusing proangiogenic factor. At this stage, the tumor reaches a maximum diameter of about 300 micrometers. The activated neovascular cells in the pre-existing vasculature respond to the pro-angiogenic factors both by chemotaxing towards higher concentrations of the proangiogenic field and by forming new blood vessels via angiogenesis. The tumor-induced neovascular network enhances oxygen supply and tumor growth. Total movie time = 75 days. Time interval between frames = 6 hours. The camera rotates at a rate of 0.1 degree per frame. Cell types: Green: normal; yellow: hypoxic; blue: necrotic; red: vascular; purple: neovascular. Axes are labeled in micrometer.(9.71 MB AVI)Click here for additional data file.


Biventricular pacing has been suggested in end-stage heart failure. We present a 59-year-old patient undergoing second re-do CABG (coronary artery bypass graft) and carotid artery endarterectomy. Ejection fraction was 15%, QRS-width 175 ms. Following the carotid and CABG procedure, an implanted single-chamber ICD (implantable cardioverter defibrillator) was upgraded to permanent biventricular DDD pacing by implantation of one epicardial left ventricular and one epicardial atrial electrode. At follow-up two months postoperatively ejection fraction had significantly improved to 45%, the patient underwent stress test with adequate load and reported a good quality of life. Widening of QRS complexes to more than 150 ms has been defined to be one independent risk factor for cardiac mortality in patients with poor left ventricular function ,3. In opFour weeks prior to the operation a 59-year-old male was admitted to hospital for unstable angina and dyspnea at rest (NYHA III-IV). Following preoperative diagnostics the patient was referred to our hospital for carotid endarterectomy and a simultaneous second re-do CABG procedure after previous surgery 7 and 15 years earlier. Concomitant diseases were diabetes, renal insufficiency and chronic hepatitis. Nine months prior to admission a single-chamber ICD system had been implanted for recurrent ventricular tachycardias. The ECG showed sinus rhythm with AV-block I° and a complete left bundle-branch block with QRS-width of 170 ms. Preoperative echocardiography demonstrated an ischemic cardiomyopathy with inferior akinesia, septum bulging and an ejection fraction of 15%, moderate mitral insufficiency II° and moderate pulmonary hypertension. Positron emission tomography revealed a globally thin and dilated left ventricle with a transmural inferior scar and hibernating myocardium in the remaining segments.Immediately before initiation of anesthesia an intra-aortic balloon pump was implanted to prevent hemodynamic instability throughout the procedure. The carotid disobliteration was followed by the coronary operation with revascularization of the left anterior descending artery and the right coronary artery using the left internal thoracic and right radial arteries. The circumflex branches were too small for revascularization. Intraoperatively the left ventricle was enlarged with an inferior scar and globally reduced contractility Following release of the aortic cross clamp and completion of the proximal aortic anastomosis, mapping of the lateral and anterior left ventricular wall was performed under echocardiographic control using an epicardial screw-in electrode CapSure Epi 4965 . Paradox septum motion disappeared with left ventricular stimulation; the most symmetric contraction was achieved by stimulating a segment at the basis of the lateral wall close to the left atrial appendage. The electrode was implanted with a pacing threshold of 1.2 V (Volt) and an R-wave sensing of 11.5 mV (milliVolt). An additional bipolar epicardial electrode Medtronic CapSure Epi 4968 was placed at the lateral wall of the right atrium. The incision at the left upper thorax was reopened; the defibrillation electrode was connected to the new three-chamber-ICD SJM Epic™ . Both epicardial electrodes were transferred into the ICD pocket through the left hemithorax and connected as well.th day he developed right-sided pneumonia and was artificially ventilated for another three days, but recovered well and could be mobilized during the following week. He was transferred to a rehabilitation centre on the 18th day with no signs of congestive heart failure . Total bypass time was 142 min. The remaining intra- and early postoperative course was uneventful except for a re-sternotomy for diffuse bleeding. The hemodynamic condition improved with a normal cardiac output throughout the intensive care stay. On the second postoperative day inotropic support was completely weaned, the intra-aortic balloon pump was removed and the patient was extubated. On the 7Two and eight months after the operation, the patient presented for a follow-up visit in good condition. He did not complain of angina or dyspnea anymore (NYHA I), his stress test up to 90 W was uneventful except for physical fatigue, no ECG changes were seen under continuous DDD pacing, QRS-width was 145 ms. Echocardiography showed an unchanged inferior akinesia, but a normal septum motion with an improved global left ventricular function. Ejection fraction was 45%, mitral insufficiency was reduced to 0-I°.Various studies have demonstrated efficacy of biventricular pacing in advanced heart failure ,3,7. TheCompared to a transvenous approach, epicardial electrode implantation allows mapping of the whole left ventricular wall with optimization of the electrode placement and adds only a short period of operation time. The positive effect of biventricular stimulation on regional wall contractility can be monitored by transesophageal echocardiography. In our case inferior wall and interventricular septum motion abnormality were demonstrated initially, the latter a phenomenon which is very common in open heart surgery not only in patients with reduced ventricular function and left bundle-branch block. Biventricular pacing led to immediate normalization of this septum bulging.During early follow-up the patient improved dramatically regarding his NYHA classification due to a combination of myocardial revascularization and permanent biventricular pacing. His improved quality of life and his almost normal stress test could be related to an increased ejection fraction and decreased mitral insufficiency.In summary, we suggest a generous consideration of permanent biventricular stimulation during open heart surgery in patients with severely impaired left ventricular function and widened QRS-complexes. The technique adds only little extra time, allows optimal lead location and monitoring by transesophageal echocardiography. It facilitates the early postoperative phase and adds extra benefit to the quality of life in these high risk patients. Long-term effects and impact on mortality during follow-up will have to be investigated.The authors declare that they have no competing interests.OD and PK performed the operation.NM analyzed the patient data.AM did the final proof-reading of the manuscript.Written informed consent was obtained from the patient for publication of this case report and accompanying images. A copy of the written consent is available for review by the Editor-in-Chief of this journal.


Assessment of visual fixation is commonly used in the clinical examination of patients with disorders of consciousness. However, different international guidelines seem to disagree whether fixation is compatible with the diagnosis of the vegetative state  or is a sufficient sign of consciousness and higher order cortical processing.We here studied cerebral metabolism in ten patients with chronic post-anoxic encephalopathy and 39 age-matched healthy controls. Five patients were in a vegetative state (without fixation) and five presented visual fixation but otherwise showed all criteria typical of the vegetative state. Patients were matched for age, etiology and time since insult and were followed by repeated Coma Recovery Scale-Revised (CRS-R) assessments for at least 1 year. Sustained visual fixation was considered as present when the eyes refixated a moving target for more than 2 seconds as defined by CRS-R criteria.Patients without fixation showed metabolic dysfunction in a widespread fronto-parietal cortical network (with only sparing of the brainstem and cerebellum) which was not different from the brain function seen in patients with visual fixation. Cortico-cortical functional connectivity with visual cortex showed no difference between both patient groups. Recovery rates did not differ between patients without or with fixation (none of the patients showed good outcome).Our findings suggest that sustained visual fixation in (non-traumatic) disorders of consciousness does not necessarily reflect consciousness and higher order cortical brain function. It is still a matter of debate whether visual fixation indicates "automatic" subcortical processing  or wh18F]-fluorodeoxyglucose-PET (FDG-PET) in ten patients with chronic (>4 weeks) post-anoxic encephalopathy  and 39 age-matched healthy controls . Patients were assessed by means of the Coma Recovery Scale-Revised (CRS-R) [Cerebral metabolic rates for glucose (CMRGlu)  were stuVisual fixation was assessed as defined in the CRS-R : a brighhttp://www.fil.ion.ucl.ac.uk/spm). We looked for brain regions where CMRGlu was different between: (i) patients without fixation and healthy controls (ii) patients with fixation and healthy controls (iii) patients without fixation as compared to patients with fixation. In a second step, we used a psychophysiological interaction analysis [FDG-PET data were pre-processed  and analanalysis  to test analysis ) in patianalysis ). ResultWhen compared to healthy participants, VS patients without fixation showed metabolic dysfunction in a widespread cerebral network encompassing bilateral thalami and fronto-temporo-parietal associative cortices. Areas that were relatively spared were confined to the brainstem and cerebellum. Patients with visual fixation but otherwise showing clinical characteristics typical of VS showed a similar pattern of metabolic dysfunction . We also point out the difficulty with non-significant findings in small cohort studies where the lack of difference may be the result of weak statistical power. However, the graphical illustration of the single-subject data of the functional segregation analysis shows an almost complete overlap between both patient groups and the functional integration analysis  shows coOur findings are in line with previous studies of "automatic" visual fixation of salient stimuli in blindsight , hemianoIn our view, the present results are of interest to clinical neurologists, who have taken visual fixation and tracking as being an important step in recovery of consciousness from the vegetative state . The herIn conclusion, our findings suggest that in (non-traumatic) disorders of consciousness, sustained visual fixation is not accompanied by higher order frontoparietal integrative cortical brain function which is assumed to be associated with conscious awareness .The authors declare that they have no competing interests.MAB made substantial contributions to acquisition of data, analysis and interpretation of data as well as in drafting the manuscript. AV, CS and MB were implied in the interpretation of data as well as in drafting the manuscript; gave critical revision of the manuscript for important intellectual content. OG and AD were implied in collecting behavioural data. SM, RH and GM were involved in drafting the manuscript. SL made substantial contributions to conception and design and supervised this study. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2377/10/35/prepub


Parametric linkage methods for quantitative trait locus mapping require explicit specification of the probability model of the quantitative trait and hence can lead to misleading linkage inferences when the model assumptions are not valid. Ghosh and Majumder developed a nonparametric regression method based on kernel-smoothing for linkage mapping of quantitative trait locus using squared differences in trait values of independent sib pairs, which is relatively more robust than parametric methods with respect to violations in distributional assumptions. In this study, we modify the above mentioned nonparametric regression method by considering local linear polynomials instead of the Nadaraya-Watson estimator and squared sums of sib-pair trait values in addition to squared differences to perform a genome-wide scan of rheumatoid factor-IgM levels on sib pairs in the Genetic Analysis Workshop 15 simulated data set. We obtain significant evidence of linkage very close to the quantitative trait locus controlling for RF-IgM. We find that the simultaneous use of squared differences and squared sums increases the power to detect linkage compared to using only squared differences. However, because of all the sib pairs are selected for rheumatoid arthritis, there is reduced variance of RF-IgM values, and empirical power to detect linkage is not very high. We also compare the performance of our method with two linear regression approaches: the classical Haseman-Elston method using squared sib-pair trait differences and its extension proposed by Elston et al. using mean-corrected sib-pair cross-products. We find that the proposed nonparametric method yields more power than the linear regression approaches. Heritable quantitative characters are precursors of a clinical end-point trait. Because end-point traits are usually binary in nature (affected/unaffected) and hence contain minimal information on variation within trait genotypes, it may be statistically more powerful to use a correlated quantitative phenotype for identifying genes for the underlying complex trait. Unlike qualitative or binary traits, which can be characterized completely by allele frequencies and genotypic penetrances, quantitative traits require a stronger layer of modeling: the probability distribution of the underlying trait. Thus, compared to likelihood-based approaches like variance components ,2, whichFor our analyses, we used data on RF-IgM levels and genome-wide information on 730 microsatellite marker loci distributed over the 22 autosomal chromosomes. Our method utilizes marker genotype data on 1500 independent sib pairs and their parents for identity-by-descent (IBD) computations. The nonparametric regressions for the linkage scan are based on the RF-IgM and IBD data. We performed our analyses on all 100 available replicates.yij denotes the RF-IgM of the jth sib in the ith family, i = 1, 2,..., 1500; j = 1, 2; and ith sib pair at an arbitrary point p on the genome. We define Ui = (yi1 - yi2)2 and Vi = (yi1 + yi2)2. The classical Haseman-Elston method [Ui values) on estimated marker IBD scores  to consider a linkage finding to be statistically significant. Since the "answers" were available to us, we considered a linkage peak to be true positive only if both the following criteria were satisfied: it was within a 20 cM window (10 cM on either side) of the true position of a QTL and all other markers within this window provided significant evidence of linkage. Hence, we have assessed the empirical power and the false-positive error rate based on the proportion of replicates yielding significant linkage peaks.We performed our nonparametric regression analyses on all 22 autosomal chromosomes for all 100 available replications. We compared the results of the nonparametric regression with those of the classical Haseman-Elston regression using squared differences  using mep-value < 0.05) at marker STRP11_22 (113 cM) on chromosome 11 in 17 replications using squared differences only and in 31 replications using both the squared differences and the squared sums. All of the other markers within the 20-cM window of the position of the QTL have also provided evidence of linkage at level 0.05 for all these replications. Although given a threshold of p < 0.001 for a linkage peak to be statistical significant, the empirical power was only 0.1 when only squared differences were used and 0.23 when both squared differences and squared sums were used, we note that the linkage peak is close to Locus F (115 cM), the QTL controlling RF-IgM. However, the major aim of the study, that is, the belief that the combined use of squared differences and squared sums is more powerful than using only the squared sums is validated by our results. We also found that there was no other marker which provided a statistical evidence of linkage at level 0.05 in more than three replications.Based on the proposed nonparametric regression, we obtained a linkage peak  declare that they have no competing interests.


In the years 1979-1982, 83 patients with malignant teratoma of the testis who had retroperitoneal adenopathy at presentation or after a period of surveillance were treated. Complete radiological resolution of disease was obtained in 34 patients and a residual mass remained in 26, the remainder having progression of the para-aortic or other metastatic disease. There was no para-aortic relapse in 47 patients receiving radiotherapy post-chemotherapy whereas 2/11 who did not receive radiotherapy or an immediate retroperitoneal node dissection relapsed. Morbidity from radiotherapy was minimal apart from subcutaneous fibrosis in the irradiated area of 6 patients. It is concluded that radiotherapy is effective in sterilising minimal residual tumour post-chemotherapy and may be considered as an alternative to surgery.


To assess comorbid DSM-IV-TR Axis I disorders in adolescent inpatients referred for treatment of substance use disorders.151 patients  were consecutively assessed with the Composite International Diagnostic Interview (CIDI) and standardized clinical questionnaires to assess mental disorders, symptom distress, psychosocial variables and detailed aspects of drug use. A consecutively referred subgroup of these 151 patients consisting of 65 underage patients  was additionally assessed with the modules for attention-deficit/hyperactivity disorder (ADHD) and conduct disorder (CD) using The Schedule for Affective Disorders and Schizophrenia for school-aged children (K-SADS-PL).128 (84.8%) of the 151 patients were dependent on at least one substance, the remaining patients fulfilled diagnostic criteria for abuse only. 40.5% of the participants fulfilled criteria for at least one comorbid present Axis I disorder other than substance use disorders . High prevalences of present mood disorder (19.2%), somatoform disorders (9.3%), and anxiety disorders (22.5%) were found. The 37 female participants showed a significantly higher risk for lifetime comorbid disorders; the gender difference was significantly pronounced for anxiety and somatoform disorders. Data from the underage subgroup revealed a high prevalence for present CD (41.5%). 33% of the 106 patients  who were within the mandatory school age had not attended school for at least a two-month period prior to admission. In addition, 51.4% had been temporarily expelled from school at least once.The present data validates previous findings of high psychiatric comorbidity in adolescent patients with substance use disorders. The high rates of school refusal and conduct disorder indicate the severity of psychosocial impairment. The misuse of psychotropic substances is one of the most prevalent mental disorders in industrial nations and drug use is a frequent problem therapists in both adolescent and adult psychiatric settings must deal with. Johnston et al.  stated tReflecting the health care system in many countries, most studies were conducted on outpatients or patients in residential programs. As a result, there is insufficient knowledge about psychiatric comorbidity in adolescent inpatients. As far as we know, only Deas et al.  and HoveTo evaluate the temporal stability and developmental pathways of comorbid mental disorders, data on both current and lifetime comorbidity are required. However, to our knowledge, all recent studies limit the timeframe to either current or lifetime disorders. Furthermore, even the rates of current disorders are not based on the same timeframe; 12-month-, six-month and point prevalences of disorders are accepted indices to describe rates of present morbidity.In light of the aforementioned limitations it should be noted that adolescent SUD patients very often suffer from externalizing disorders  and to a somewhat lesser extent from anxiety and mood disorders. Based on ten recent studies, Couwenbergh et al.  computedLittle research has been conducted on the consequences of maladaptive substance use concerning, school refusal and the link to comorbid mental disorders. Although some researchers ,27,29 dePsychiatric SUD treatment of adolescent inpatients differs in various ways from SUD treatment or detoxification of adults. Many practitioners agree that inpatient adolescent SUD treatment far more often has to account for specific difficulties like inactivity, high rates of treatment dropout and oppositional disorders. In many cases it remains unclear whether these problems are part of an age-appropriate developmental process or symptoms of a mental disorder. In the case of a comorbid axis-I disorder, misinterpreting these symptoms as normal adolescent-like behaviour or part of the substance use disorder would possibly delay the treatment of the comorbid disorder for a considerable time.Although some practice-oriented treatment programs have been developed in the last decade many therapy concepts focus on consumption-related symptoms of SUD like withdrawal or maintenance of abstinence. Relating to the detoxification of adults or outpatient treatment of moderate SUD, this priority may be a reasonable approach. In the area of inpatient SUD treatment of adolescents this procedure runs the risk of neglecting severe psychosocial symptoms like school refusal or evolving delinquent/aggressive behaviour. This present study aims to provide further comprehensive data on psychiatric comorbidity of adolescents with substance use disorders with an additional focus on both gender aspects and school refusal. Furthermore we address some developmental psychopathological data as we include both lifetime and present axis-I diagnoses considering the changes in psychopathology.Participants were 151  adolescents and young adults (≤22 years) referred for inpatient substance abuse treatment between April 2005 and December 2006. Patients were consecutively recruited in SUD-treatment units of the Rheinische Kliniken Essen (99 patients) and Kreiskrankenhaus Gummersbach - Klinik Marienheide (52 patients). Both units are located within child and adolescent psychiatric departments, providing full-service psychiatric health care. The Rheinische Kliniken Essen is situated in a metropolitan area of Germany whereas the Klinik Marienheide is located in a rural region. The procedure of admission to both drug-specific inpatient programs was comparable; in both units patients were required to be heavy drug users with clinically significant impairment or distress. Inclusion criteria were at least one SUD (other than tobacco-SUD) according to DSM-IV-TR , age betDuring the second or third week of inpatient treatment, independent face-to-face interviews and questionnaires were conducted with the subjects. All interviews and questionnaires were administered by trained medical students or graduated, experienced clinical psychologists. One experienced clinical psychologist for each hospital acted as supervisor and guided the examiners. The clinical examinations lasted three and a half hours on average and were composed of six modules.(1) The German edition ,36 of th(2) To access the DSM-IV-TR disorders ADHD and conduct disorder (DSM-IV-TR code 312.8), which are not included in the CIDI, the corresponding modules of the Schedule for Affective Disorders and schizophrenia for school-aged children - Present and Lifetime Version - German version  -40 were (3) The Fagerström Test for Nicotine Dependence (FTND)  was used(4) The German version  of the S(5) Detailed information about drug-consumption for all relevant substances  obtained by comprehensive semi-structured interviews was recorded.(6) A semi-structured interview slightly modified according to the Adolescent Drug Abuse Diagnosis (ADAD)  was usedDue to the fact that some parents of the participants either did not cooperate in a required manner or had no contact to their children for a long time, all interviews and questionnaires were carried out with the patients only.Means, standard deviations and percentages were calculated to describe aspects of drug-use. To study possible differences between groups, the phi coefficient was used to examine nominal data, Student's t-test for interval and ANOVA for comparisons of interval data with more than two groups. Tests of significance were two-tailed using exact tests procedure for nonparametric statistics. The level of statistical significance was set at p < .05. Missing data (in five cases) have been substituted by the mean of the respective variable. All statistical analyses were carried out using SPSS V14.0.Tobacco 99.3%), cannabis 84.8%) and alcohol (64.9%) were the most commonly used substances as well as the substances most often associated with SUD table . Regardi%, cannab.8% and apresent somatoform disorders), no statistical relationship between age and specific comorbid axis-I disorders could be found  still required mandatory schooling during the current school year upon admission. Of this subgroup, 33.0% had not at all attended school during the last two months prior to admission. The mean number of absent days for actually school attending participants (n = 71) during the last two months (46 days of school attendance) was 17.72 days (SD = 18.40). 51.4% of the school aged participants had been temporarily expelled from school at least once, 32.4% had to change schools as a disciplinary action. All participants were asked to rate their performance at school during the last year (or last year of school attendance in case of no current school attendance) on a three-point Likert scale ranging from below average (1) over average (2) to above average (3). 51.3% rated their school achievement below average, 45.3% average and 3.3% above average.Female participants suffered significantly more often from one or more lifetime and one or more present comorbid mental disorders  . In detail, female participants significantly more often fulfilled criteria for lifetime and present PTSD , present  and lifetime  anxiety disorders, present  and lifetime  somatoform disorders than males. A female preponderance (diagnoses include ADHD and CD) was also detectable in the underage subgroup but did not reach statistical significance . No significant difference in the mean number of comorbid diagnoses of patients with at least one comorbid disorder (without ADHD & CD) between females and males was found . Additional t-tests showed no significant differences in symptom distress measured with SCL-90-R between male and female participants. Data from the subgroup  indicated no relationship between gender and rates of CD or ADHD (present and lifetime).Due to the different forms of treatment, the evaluation of SUD prevalences in clinical samples is difficult. Nevertheless our results are basically consistent with other results ,22,28. IOur SUD-patients most frequently suffered from comorbid mental disorders, predominantly conduct disorder and often anxiety and mood disorders. The high general risk of present comorbidity  found in this study is comparable to rates reported by most other studies (61% to 88%) of clinical SUD-samples . In accoIn comparison with studies that assessed axis-I disorder rates in the German general population, our rates of lifetime diagnoses seem to be only slightly higher than rates found in representative cohorts: Essau et al.  scanned Except for the positive distress index in patients with at least one comorbid diagnosis, data obtained via the SCL-90-R demonstrated no clinically significant (T ≥ 60) degree of psychological distress either in patients with or without comorbidity. Considering the impairments which are most often associated with mental disorders, SUD and broken home situations, these results are difficult to interpret. Dissimulation to avoid long-term treatment, distorted self-perception and the relieving influence of inpatient treatment ("honeymoon effect") could possibly account for these results.Comparable to Hovens et al.   but led to a limited reliability of the diagnoses of CD and ADHD. Symptoms of external disorders (e.g. CD and ADHD) are underreported by adolescents in comparison to their parents . It is iThe high rate of comorbid psychopathology in inpatient SUD-patients, particularly conduct disorder has implications for therapy and framework of specialized treatment-units. Three-quarter of all patients show distinct comorbid psychopathology and SUD therapists should be able to take up this challenge. Patients with such a high rate of conduct disorder require specialised forms of treatment able to cope with high levels of aggression and treatment abortion often associated with CD.Future research should investigate the causal and temporal relationship between conduct disorder and SUD, especially in respect of early developmental trajectories. Besides mental disorders, the high rate of school refusal and truancy should also be considered as important part of the substance use problem. Existing school refusal treatment programmes should be aware of the high co-occurrence whereas SUD-treatment units should carefully evaluate psychological causes of school refusal and emphasize school reintegration. Finally, controlled longitudinal comparative studies are needed to test the possible positive effect of comorbidity-considering treatment programmes.The authors declare that they have no competing interests.Authors TL, NQ, NS, PM and JH designed the study and wrote the protocol. TL and NS conducted literature searches and provided summaries of previous research studies. TL conducted the statistical analysis. TL, AS, EO and GW conducted the assessment of the participants. TL and JH wrote the manuscript and all authors contributed to and have approved the final manuscript.All authors have read and approved the final manuscript.


Hypoxia was assessed in three murine tumour models in vivo by measuring the incorporation of 7-(4'-(2-nitroimidazole-1-yl)-butyl)-theophylline (NITP), an immunologically identifiable hypoxia marker that binds bioreductively to cells under low-oxygen conditions. Proliferating cells were labelled in the same tumours by administering the thymidine analogue bromodeoxyuridine (BrdUrd). The relative hypoxia in each cell cycle phase of cells isolated from tumours was assessed by addition of propidium iodide with analysis by flow cytometry. There was no relationship between tumour volume and hypoxia in either the anaplastic sarcoma SaF or the poorly differentiated carcinoma CaNT and only a slight negative correlation in moderately well-differentiated carcinoma Rh. The G1/G0 phase contained the greatest number of aneuploid hypoxic cells , although there were significant amounts of hypoxia present in S- and G2/M phases for all three tumours examined. However, the highest proportion of hypoxia occurred in the G2/M phase, in which up to 60% of the cells were hypoxic. Simultaneous measurement of hypoxia, proliferation and DNA content using a novel triple-staining flow cytometry method showed that hypoxic cells could actively participate in the cell cycle. In addition, the cell cycle distribution of NITP and BrdUrd labelling showed that hypoxic cells could progress through the cell cycle, although their rate of progression was slower than that of better oxygenated cells.


Primary breast carcinomas (23) with axillary-node metastases that also showed tumour cells in the efferent nodal vessels, tended to be of higher histological grade than those (21) without efferent vascular invasion. Nuclear hyperchromatism and mitosis is the factor of importance to grading in this respect. This factor also differentiated between RE+ and RE- carcinomas in this material.


This paper addresses the diagnosis and management of superior vena cava syndrome(SVCS) due to malignant intrathoracic tumours. Diagnosis of SVCS is usually establishedby bedside examination. Chest X-ray and computed tomography may be helpful, but thecavography remains the “gold-standard”. Other imaging techniques  are more of scientific interest. Bronchoscopy helps to evaluate the risk of pulmonarycomplications and endoscopic procedures often lead to histological findings. In thetreatment of malignant SVCS surgery, radiotherapy, and chemotherapy have been successfullyused. The placement of a vascular stent might be an additional or alternative possibility.There are no conclusive indication criteria and no conclusive regimen concerningpost-stenting anticoagulation. From all reported results and published papers we draw theconclusion that the immediate effects of stent implantation and the long-term results oftumour-specific therapy are complementary to one another. The stent dilates the localvenous stenosis while tumour-specific therapy has a general effect on the vascular andrespiratory situation in a multi-therapy concept.


TNF-alpha is a critical cytokine produced by Th1 cells while altered T helper 1 (Th1)-Th2 balance is found crucial for a successful pregnancy.A cohort of 132 Southern Chinese Han RSA patients and 152 controls constituted the subjects of this study. Two functional polymorphisms -308 and -238 of TNF-alpha were studied by association analysis.lack of association was found in TNF-alpha -308 SNP yet a significant difference was discovered in -238 polymorphism.This study suggested that TNF-alpha may be a risk factor in Chinese RSA patients. However the ethnic differences may also contribute to the results. Recurrent spontaneous abortion (RSA) is defined as three or more consecutive pregnancy losses before 22 gestational weeks or the spontaneous abortion of an embryo/fetus weighing less than 500 g and affecting an estimated 1 of every 100 couples preparing to have children . TTo our knowledge, this work firstly established an association between TNF-alpha gene polymorphisms and the development of URSA in Chinese population.TNF-alpha gene possibly not be a potential risk factor of URSA developing, but patients with URSA has a 3.95-fold higher frequency of -238 GG genotype adjusted by age, which demonstrated the association between -238 polymorphism and the URSA developing in Chinese population.In conclusion, the present study indicated that -308 polymorphism of The authors declare that they have no competing interests.CL collected all samples and performed the clinical tests. JW carried out the molecular genetic studies and performed the statistical analysis. SZ drafted the manuscript. BW participated in its design and coordination and helped to draft the manuscript. XM participated in the design of the study. All authors read and approved the final manuscript.


In past number of methods have been developed for predicting subcellular location of eukaryotic, prokaryotic (Gram-negative and Gram-positive bacteria) and human proteins but no method has been developed for mycobacterial proteins which may represent repertoire of potent immunogens of this dreaded pathogen. In this study, attempt has been made to develop method for predicting subcellular location of mycobacterial proteins.The models were trained and tested on 852 mycobacterial proteins and evaluated using five-fold cross-validation technique. First SVM (Support Vector Machine) model was developed using amino acid composition and overall accuracy of 82.51% was achieved with average accuracy (mean of class-wise accuracy) of 68.47%. In order to utilize evolutionary information, a SVM model was developed using PSSM (Position-Specific Scoring Matrix) profiles obtained from PSI-BLAST (Position-Specific Iterated BLAST) and overall accuracy achieved was of 86.62% with average accuracy of 73.71%. In addition, HMM (Hidden Markov Model), MEME/MAST (Multiple Em for Motif Elicitation/Motif Alignment and Search Tool) and hybrid model that combined two or more models were also developed. We achieved maximum overall accuracy of 86.8% with average accuracy of 89.00% using combination of PSSM based SVM model and MEME/MAST. Performance of our method was compared with that of the existing methods developed for predicting subcellular locations of Gram-positive bacterial proteins.A highly accurate method has been developed for predicting subcellular location of mycobacterial proteins. This method also predicts very important class of proteins that is membrane-attached proteins. This method will be useful in annotating newly sequenced or hypothetical mycobacterial proteins. Based on above study, a freely accessible web server TBpred http://www.imtech.res.in/raghava/tbpred/ has been developed. Mycobacterium tuberculosis H37Rv are yet to be assigned [According to the GOLD (Genomes OnLine Database) database  as on 12assigned . Thus itMycobacterium tuberculosis genome using experimental and predicted information [In last few years several subcellular localization prediction systems have been developed using various features of a protein like composition of amino acid, pseudo amino acid, dipeptide and Physico-chemical properties -9. Recenormation ,35.To the best of authors' knowledge no method has been developed for predicting subcellular localization of mycobacterial proteins, which has different cell wall composition than Gram-negative or Gram-positive bacteria. In this study we describe models developed for predicting four subcellular locations of mycobacterial proteins, namely cytoplasmic, Integral membrane, secretory and membrane-attached proteins ,37. A syBiologists routinely use the BLAST for similarity search; it will be interesting to know the performance of BLAST on the same dataset and cross-validation used in this study. This is important because users wish to know the advantage of any sophisticated method over routinely used method like BLAST. Thus we evaluated performance of BLAST on our dataset using five-fold cross validation where proteins in test dataset were searched using BLAST against proteins of training dataset. As shown in Table The performance of all the modules developed in this study was evaluated using 5-fold cross-validation technique. The performances of different SVM modules have been summarized in Table In this study, HMM based models have been developed for each subcellular location. The performance of HMM models for each class/location is shown in Table The HMM based method allows performing similarity search at sequence level but not at motif level. Thus in this study motifs were extracted and searched using MEME/MAST software. As shown in Table As shown above in Table In order to provide confidence in prediction, we computed reliability index (RI). It is a measure of level of certainty in a prediction. Figure It is important to compare newly developed method with existing methods. This is the first subcellular location prediction method for mycobacterial proteins, thus it is difficult to compare this method with existing methods. It is known that mycobacterium species are significantly similar to Gram-positive bacteria, thus we compare our method with existing methods developed for Gram-positive bacteria. One to one comparison of existing method with our method was not possible because number of subcellular locations predicted by these methods was different than locations predicted by our method. For example none of the existing methods predicts membrane-attached proteins. In order to provide comprehensive comparison, existing methods have been evaluated on the dataset used in this study and presented by confusion matrix Table . First phttp://www.imtech.res.in/raghava/tbpred/.Various SVM modules developed in the present study were implemented into a web server, TBpred, for predicting the subcellular localization of mycobacterial proteins. User can select from amino acid composition, dipeptide composition and PSSM based SVM models or a hybrid model for prediction. The common gateway interface (CGI) script for TBpred was written using PERL 5.03. This server is installed on a Sun Server (420E) under a UNIX (Solaris 7) environment. TBpred is freely available at Several methods have been developed for predicting subcellular location of eukaryotic, prokaryotic (Gram-negative bacteria) and human proteins but no method is available for mycobacterial proteins. Thus there was a need to develop a dedicated method for predicting subcellular localization of mycobacterial proteins. There are two reasons for developing subcellular localization method specially for mycobacterial proteins; i) organism specific subcellular localization method(s) performs better than generalized methods ,27-29; iIn order to overcome these limitations we developed HMM based models for predicting subcellular location. The performance of HMM based model was reasonable for secretory and membrane-attached proteins but its performance was poor for other two classes Table . It seemA new subcellular class of mycobacterial proteins named "membrane-attached by lipid anchor" has been introduced for the first time. This class of protein may play a role in enhancing the immune response of the host by acting as surface antigens. Thus the search for a potential vaccine/drug target for this immensely important bacterial pathogen by the experimental researchers will greatly be appended by the prediction algorithm developed in this study. Moreover, the comparison of TBpred prediction efficiency with existing methods developed for Gram- positive bacteria supported our earlier assumption that organism specific classifier performs better than the generalised one.The mycobacterial protein sequences were extracted from release 48 of Swiss-Prot. Initially, we got 1365 mycobacterial proteins; after removing the fragments and the non-experimental qualifier "by similarity" we got 882 proteins. These 882 proteins belong to 13 subcellular locations as shown in Table Ideally one should remove similar proteins from data in order to generate the non-redundant data, as similar protein in training and testing data influence the performance of a method. We computed level of similarity in proteins belonging to a subcellular location using CD-HIT. As shown in Table Ideally one should evaluate newly developed method using jack-knife method  ,45. In jlight, which is widely used for developing methods in the field of bioinformatics [light binary classifier using 1-vs-r (one-versus-rest) approach, for developing model for predicting multiple locations. In this 1-vs-r approach a SVM model was built for each class by considering proteins of that class positive and proteins of rest of the classes as negative.In this study, Support Vector Machine has been implemented using SVMormatics ,45-51. WThe percent amino acid composition of each amino acid was calculated using standard formula described in the past . These cThe PSSM profile for each protein was generated using PSI-BLAST  by searcHMM turns a multiple sequence alignment into a position specific searching system suitable for searching databases for remotely homologous sequences. HMM analysis complements standard pair wise comparison method for large-scale sequence analysis . HMM proMotif is a pattern of nucleotides or amino acids that appear in a DNA or protein family. The MEME/MAST consists of two programs, one allows discovery of motifs shared by closely related sequences (MEME)  and the We combined the output of MEME/MAST and the output SVM module. Firstly, a comprehensive list was generated that encapsulate the hits, along with their corresponding E-value, from all MEME/MAST model  and the SVM prediction for each protein in the dataset. The MEME/MAST decision (if any) is given priority upon SVM prediction in the final assignment of class to a particular protein sample. In case if all MEME/MAST models generated hits for a sample, the sample was classified into the model generating hit with lowest E-value. Moreover if the lowest E-value is shared by more than one model , the final decision was taken on consensus among MEME/MAST and SVM models. If MAST produces no hit at given E-value then SVM model was used to predict subcellular location of a protein.The performance of all modules developed in this study was computed using standard parameters like accuracy (Acc) and Matthews correlation coefficient (MCC). Following equations were used to compute these parameters -x can be any subcellular location , exp(x) is the number of sequences observed in location x, p(x) is the number of correctly predicted sequences of location x, n(x) is the number of correctly predicted sequences not of location x, u(x) is the number of under-predicted sequences and o(x) is the number of over-predicted sequences.Where In this study we computed both overall and average accuracy. The overall accuracy is the percent of correctly predicted proteins irrespective of class. The average accuracy is mean accuracy of four classes. Both type of accuracy have their advantage and disadvantage.Reliability index is a simple indication of level of certainty in the prediction. The strategy followed to calculate the RI is similar to that mentioned by .Assignment of RI to each sequence is based upon the difference of highest and the second highest scores of various 1-v-r SVMs in the multi-class classification. RI is defined as:MR and SS created datasets, developed various modules and evaluated all modules. GPSR conceived the idea, coordinated it and refined the manuscript drafted by MR and SS. All the authors have read and approved the final manuscript.Supplementary material. It consists of various tables and figures that were somehow supportive to the conducted study.Click here for file


We have shown previously that exercise benefits stress resistance and stress coping capabilities. Furthermore, we reported recently that epigenetic changes related to gene transcription are involved in memory formation of stressful events. In view of the enhanced coping capabilities in exercised subjects we investigated epigenetic, gene expression and behavioral changes in 4-weeks voluntarily exercised rats.Exercised and control rats coped differently when exposed to a novel environment. Whereas the control rats explored the new cage for the complete 30-min period, exercised animals only did so during the first 15 min after which they returned to sleeping or resting behavior. Both groups of animals showed similar behavioral responses in the initial forced swim session. When re-tested 24 h later however the exercised rats showed significantly more immobility behavior and less struggling and swimming. If rats were killed at 2 h after novelty or the initial swim test, i.e. at the peak of histone H3 phospho-acetylation and c-Fos induction, then the exercised rats showed a significantly higher number of dentate granule neurons expressing the histone modifications and immediate-early gene induction.Thus, irrespective of the behavioral response in the novel cage or initial forced swim session, the impact of the event at the dentate gyrus level was greater in exercised rats than in control animals. Furthermore, in view of our concept that the neuronal response in the dentate gyrus after forced swimming is involved in memory formation of the stressful event, the observations in exercised rats of enhanced neuronal responses as well as higher immobility responses in the re-test are consistent with the reportedly improved cognitive performance in these animals. Thus, improved stress coping in exercised subjects seems to involve enhanced cognitive capabilities possibly resulting from distinct epigenetic mechanisms in dentate gyrus neurons. It is now well established that regular physical exercise has a positive impact on a range of biological systems, including the brain In particular, we investigated the role of epigenetic mechanisms in the brain involved in transcriptional activation in coordinating adaptive behavioral responses to stressful events. Epigenetic mechanisms comprise of post-translational modifications of DNA and histone proteins within the chromatin structure, such as the methylation of DNA, and the acetylation, methylation, phosphorylation and other modifications of the N-terminal tails of distinct histone molecules in vitro work has indeed shown that phospho-acetylation of histone H3 of the promotor region of the c-Fos gene occurs at induction of this immediate-early gene Indeed, we could show that psychologically stressful events such as forced swimming and exposure to a novel environment enhances the phosphorylation and phospho-acetylation of histone H3 in a distinct population of dentate gyrus granule neurons in the hippocampus Here we investigated whether changes in histone H3 phospho-acetylation and gene expression responses would be involved in the enhanced stress coping capabilities seen in exercised subjects. Therefore, we subjected exercised and control rats to novelty exposure and forced swimming and investigated changes in dentate gyrus histone H3 phospho-acetylation and c-Fos expression, and acute behavioral responses as well as memory formation of the event.Male Sprague-Dawley rats  were singly housed under standard lighting (14:10-hour light/dark cycle), humidity (50–60%) and temperature (22–23°C) conditions. Food and water were available ad libitum.After habituation to the housing conditions for 5 days, the experimental group was allowed free access to a running wheel (diameter 34 cm) in their home cages for a period of four weeks. The rats ran approximately 4–7 km per night which is in agreement with other reports All experiments were carried out four weeks after voluntary exercise (or non-exercise) and between 8:00 and 12:00 h. For killing, individual rats were quickly anaesthetized (<15 sec) in a glass jar containing isoflurane  vapor, after which animals were decapitated immediately and their whole brains removed, snap frozen in isopentane at −40°C and deep-frozen in dry ice. Brains were stored at −80°C.To induce novelty stress, as reported before For forced swimming, as reported before For determination of forced swimming-induced acquisition of behavioral immobility, separate groups of rats were forced to swim for 15 min (as described above) and 24 h later were subjected again to forced swimming, in this case for 5 min . Behavioral immobility (or floating) is a behavioral state in which the animal retains an immobile posture displaying only enough movement to keep the head above water. Behavior in the initial test and re-test was recorded as described above and scored at a later time point. Three distinct behaviors were scored  every 10 s for the entire duration of the test and the re-test. In all cases scoring was conducted in a blinded fashion.Brain tissues were cut into coronal sections using a cryostat and mounted on glass slides  previously coated with poly-L-lysine (Sigma). Sections of rat brain were taken from the nucleus accumbens, PVN and the dorsal hippocampus in accordance with the atlas of Paxinos and Watson (1986) . Sections were stored at −20°C until use. Storage at this temperature does not affect levels of P(Ser10)-Ac(Lys14)-H3 or c-fos in sections.2O2. To improve antibody penetration of the tissue, the sections were incubated for 1 h in 0.2% Triton X-100, followed by blocking of sections with 5% goat serum in PBS to prevent non-specific binding. The primary antibodies were diluted in 1.5% goat serum/PBS. Rabbit polyclonal antibody against P(Ser10)-Ac(Lys14)-H3  was purchased from Upstate  and the rabbit anti-c-Fos antibody  was purchased from Calbiochem . Incubation with primary antibodies occurred overnight at room temperature.Immunohistochemical staining using diaminobenzidine (DAB) was conducted according to standard protocol as previously described + substrate  for the development of immunostaining were used according to company instructions. After dehydration in ethanol, sections were finally mounted using Histomount (Fisher) and coverslipped.Biotinylated secondary antibody, avidin-biotin-peroxidase complex and DAB/Ni+ neurons and c-Fos-positive (c-Fos+) neurons in the dentate gyrus  were counted by an individual blind to the treatment. The location of each positive neuron was distinguished between the dorsal and ventral blade of the dentate gyrus. For analysis of the nucleus accumbens, two sample areas dorso-lateral to the anterior commissure were used to count positive neurons. For each antibody and brain region at least 2 assays were performed. The assays provided similar results and data of one assay is presented here. The experimental data were statistically evaluated using ANOVA and, if significant, followed by the post-hoc Bonferroni test as appropriate. Behavioral data of the novelty challenge were statistically tested using ANOVA with repeated measures followed by Student's t-test in appropriate cases. Forced swim test data were evaluated using Student's t-test. The experimental data were considered to be statistically different from control data when P<0.05.The numbers of both P(Ser10)-Ac(Lys14)-H3+ neurons were found elsewhere in the brain. As previous in vitroin vivo+ and c-Fos+ neurons in the dentate gyrus was sparse which concurs with previous observations + and c-Fos+ neurons .In terms of P(Ser10)-Ac(Lys14)-H3 animals . If the  animals . However animals . Apparen+ neurons in the dentate gyrus of exercised animals  as well as behavioral responses upon re-exposure to the forced swim challenge (i.e. the re-test). Previously, we have shown that histone H3 phospho-acetylation and c-Fos induction in dentate granule neurons is required for the acquisition of immobility behavior seen in the re-test + neurons after forced swimming was significantly higher in the exercised rats than in the control animals -Ac(Lys14)-H3, at least if the whole dentate gyrus was considered . Thus, fBehavior of rats during the initial forced swim test and the re-test were recorded and scored to assess whether changes in histone H3 phospho-acetylation and c-Fos expression in the exercised rats were related to behavioral changes in this paradigm. In the initial forced swim test, control and exercised rats showed largely similar behaviors , with si+ neurons in an area of the nucleus accumbens showing highest numbers of immuno-positive neurons, comprising parts of both the core and shell region. Forced swimming induced a marked increase in the number of c-Fos+ neurons in the nucleus accumbens of both control and exercised animals. The increase, however, was similar in both groups , a stress-sensitive nucleus that plays a principal role in hypothalamic-pituitary-adrenocortical (HPA) axis regulation In the present study, we show that exercised rats present improved coping responses and memory performance after exposure to a novel environment or a forced swim test. Furthermore, these behavioral changes were associated with enhanced responses in histone H3 phospho-acetylation and c-Fos in dentate gyrus granule neurons. In contrast, the immediate early gene responses in the nucleus accumbens and PVN were similar in exercised and sedentary control animals. Therefore, it appears that changes in histone H3 phospho-acetylation and gene expression responses in the dentate gyrus are involved in the enhanced stress-coping capabilities seen in exercised animals.Exercised rats adopted a different coping strategy than control animals when faced with the novelty challenge. They exhibited less emotionality than their non-running counterparts, exploring their new environment initially but then settling down. In contrast, the non-running, sedentary rats remained active virtually for the full 30 min. These findings correspond with our previous work in which we showed that regular voluntary exercise reduces anxiety-related behavior and novelty-induced glucocorticoid responses Although the novelty situation appeared to have less influence on the exercised rats, they showed significantly higher dentate histone H3 phospho-acetylation and c-Fos responses than the control animals. In both groups of rats the challenge mainly impacted on the dorsal  blade of the dentate gyrus which is in agreement with our previous results + and c-Fos+ neurons in the dentate gyrus. In conjunction with observations in the novelty paradigm, it seems that the neuronal response does not directly relate to the immediate behavioral reaction during the challenge but rather relates to how the information is being processed by the dentate gyrus in the hours after the challenge. The difference in processing materialized the next day when animals were re-exposed to the forced swim test. At this time, the exercised rats showed more pronounced immobility behavior than the sedentary controls. We reported before that the phospho-acetylation of histone H3 and c-Fos induction in dentate granule cells seen after the initial forced swim test is strongly associated with the immobility behavior response observed 24 h later in the re-test In contrast to the novelty challenge test, the forced swim test is a test to address changes in active coping styles such as struggling and swimming as well as adaptive coping strategies such as immobility or floating behavior. Apart from the moderate increase in swimming behavior during the initial test in exercised rats, there were no significant differences between the control and exercised animals. Nevertheless, there were substantial differences regarding the impact of the challenge on dentate gyrus granule neurons. After forced swimming, the exercised rats, as compared to the sedentary animals, showed a significantly higher number of P(Ser10)-Ac(Lys14)-H3Our previous work has shown that the phospho-acetylation of histone H3 in dentate neurons after forced swimming and novelty is brought about by concurrent signaling via the GR and NMDA/ERK/MSK pathways Here we showed that exercised rats have improved capabilities to cope with psychologically stressful challenges. This enhanced stress coping materialized during the exposure to a novel environment and when re-submitted to a forced swim challenge. This improved adaptive capacity may be the logical consequence of the complex of elevated cognitive abilities, lowered anxiety levels and decreased impulsiveness known of exercised subjects