Ttr gene coding transthyretin involved in amyloidosis, seizures, stroke-like episodes, or dementia. Strain-specific HA transcriptome affected by CMS was associated with deregulation of genes involved in insulin secretion , neuropeptide hormone activity (Nts and Trh), and dopamine receptor mediated signaling pathway . LA transcriptome affected by CMS was associated with genes involved in behavioral response to stimulus , immune effector processes , diacylglycerol binding , and long-term depression  and/or coding elements of dendrites  and myelin proteins . The results indicate significant contribution of genetic background to differences in stress response gene expression in the mouse prefrontal cortex.There is increasing evidence that depression derives from the impact of environmental pressure on genetically susceptible individuals. We analyzed the effects of chronic mild stress (CMS) on prefrontal cortex transcriptome of two strains of mice bred for high (HA)and low (LA) swim stress-induced analgesia that differ in basal transcriptomic profiles and depression-like behaviors. We found that CMS affected 96 and 92 genes in HA and LA mice, respectively. Among genes with the same expression pattern in both strains after CMS, we observed robust upregulation of The online version of this article (doi:10.1007/s12031-012-9850-1) contains supplementary material, which is available to authorized users. CMS was adapted from the procedures developed by Willner et al.  according to the manufacturer's protocol and starting with 100 ng total RNA. Samples were purified with the RNeasy kit  according to the manufacturer's protocol.Hybridization to the Sentrix MouseRef-8 Expression BeadChip , washing, and scanning were performed according to the Illumina BeadStation 500× manual. Sentrix MouseRef-8 Expression BeadChip contains approximately 24,000 well-annotated RefSeq 50-mer oligonucleotide probes per array. Data were extracted using software provided by the manufacturer. Illumina Beadstudio v2 software with the default settings for gene expression analysis was used.www.bioconductor.org). Data preprocessing step involved normalization of expression levels with quantile method was preceded by log2 transformation. Linear model fitting was performed for the pre-processed dataset. The empirical Bayes analysis was performed in order to identify differentially expressed genes by testing whether the contrast coefficients from the linear models can be assumed equal zero. Genes considered to be significantly differentially expressed with logged fold-change are greater than 0.5 and adjusted p value is less than 0.05. Benjamini and Hochberg method . EASE takes into account the frequencies of genes belonging to particular Gene Ontology terms  among the genes found to be regulated and among all genes studied in the experiment. EASE performs statistical analysis to detect overrepresented functional gene categories in the data set compared with all genes on the arrays. GO terms are reported with corresponding EASE scores—it is a conservative statistical test that gives the upper bound of the distribution of the Jackknife Fisher exact probabilities and favors robust categories. Functional gene categories were considered significantly overrepresented at p < 0.05. Genes to functional categories were classified with biological process, molecular function, and cellular component ontologies.Gene lists (GenBank accession numbers) from microarray results were submitted to the Expression Analysis Systemic Explorer . In KEGG database, distributions of the differentially expressed genes were classified among biochemical pathways. Microarray data from a single gene in a pathway do not suffice to describe a regulatory mechanism of the pathway. Therefore, only pathways with the microarray information for at least two genes were considered. To identify significantly overrepresented biological categories and KEGG pathways within the lists of differentially expressed genes, the threshold of EASE Score for the enrichment analysis was set at p ≤ 0.05.Analysis of the genes' association with physiological pathways was performed using the Kyoto Encyclopedia of Genes and Genomes database . Biologically meaningful clusters consist of genes that are annotated into specific functional annotation groups. Grouping genes based on functional similarity can systematically enhance biological interpretation of large lists of genes derived from high throughput studies. The Functional Classification Tool generates a gene-to-gene similarity matrix based on shared functional annotation using over 75,000 terms from 14 functional annotation sources. DAVID clustering algorithms classifies highly related genes into functionally related groups. The classification stringency of the cluster analysis was set at “high” level. To avoid over-counting duplicated genes in the chart report view, the Fisher exact statistics was calculated based on corresponding DAVID gene IDs by which all redundancies in original IDs are removed.For a more insightful view of the relationships between annotation categories compared with chart, genes were clustered in the DAVID 6.7 Functional Annotation Clustering module (n = 15) per experimental group RNA samples, which were used in the microarray experiment. For reference, two housekeeping genes, selected from ten commonly used reference genes using previous methodology  and hypoxanthine guanine phosphoribosyl transferase 1 (Hprt1). Primers were designed using ExonPrimer software (http://ihg2.helmholtz-muenchen.de/ihg/ExonPrimer.html)  using Mus musculus GenBank sequences. All primers produced amplicons which spanned two exons each in highly conserved coding regions and included all known alternatively spliced mRNA variantsTo validate the results of microarrays, quantitative real-time RT-PCR (qPCR) with SYBR Green technique was performed as described previously  on the transcriptional effects of CMS in prefrontal cortex. Gene analysis was performed by one color hybridization of the 24 K microarray in HA and LA, naïve and CMS mice. Three independent biological replicates of microarray were prepared for each group of mice. To minimize the influence of individual differences between the animals and variation introduced by dissection and tissue preparation, total RNA separately isolated from several PFCs was pooled. Each pool, containing total RNA from five individuals, was separately converted to cRNA and hybridized to a single microarray.http://www.biogps.gnf.org/). Cell-type classification analysis of differentially expressed genes, according to Cahoy (http://www.genecards.org/) screening and Ingenuity Pathway Analysis (IPA) (http://www.ingenuity.com/) of top canonical pathways, revealed that most of them are characteristic for neurons and oligodendrocytes.The identified genes are considered to be expressed in the prefrontal cortex according to Novartis Gene Expression Atlas  Table S. Genes annotated to these probe sets were considered to be expressed in the examined brain tissue according to Novartis Gene Expression Atlas (www.biogps.gnf.org).Comparison of basal gene expression profiles between the HA and LA strains identified 193 transcripts with different levels of mRNA in the PFC that met the criteria of logged fold change greater than 0.5 and p < 0.05) over-expressed biological processes terms associated with the up- and downregulated genes between strains were found with the DAVID Functional Annotation Tool. We identified functional categories of biological process , neuron projection , cell communication , synaptic transmission , lipid binding , and calcium ion binding  , response to stimulus , structural constituent of ribosome , and coding cellular components such as cytosol and cytoplasmic parts , cell cortex parts , macromolecular complex , secretory granule and cytoplasmic membrane-bounded vesicle , and ribosome  , six genes were classified into signal transduction and intracellular signaling cascade cluster , three genes into ion binding cluster , and three genes into transport and establishment and localization cluster   over-expressed terms associated with the up- and downregulated genes were found in both strains subjected to CMS. Functional annotation was done in each strain on a list of up- and downregulated transcripts separately. In HA mice, CMS affected the upregulation of genes involved in, e.g., intracellular signaling, ion binding, neuropeptide hormone activity, and metabolism of cAMP and nucleotides .To confirm differential expression indicated by the microarray expression patterns, the quantitative real-time reverse transcription PCR was used (qPCR). qPCR were performed using aliquots of the non-pooled total RNA. Following genes were selected from the list of the significant functional clusters in each strain: Environmental factors and genes contribute to the manifestation of phenotype which, however, may occur or not. The proper activity of a complex organ such as the brain relies on precise temporal and spatial gene expression patterns. The phenotype is a result of a cascade of transcriptional events that finally triggers gene expression and determines the function of the particular brain structures. While a certain set of genes is constantly expressed to maintain the organ structure, a varying number of genes are regulated according to the metabolic demand of the surrounding organism. PFC is the brain region that is most sensitive to the detrimental effects of stress exposure. Abilities of PFC depend on proper neuronal network connections, which are highly sensitive to their neurochemical environment Arnsten . Since eIt is important to have animal models of stress response to understand the mechanisms that render individuals vulnerable. In this study, we have focused on the comparison of prefrontal cortex transcription profiles between naive and stressed inbred mice strains with different sensitivity to stress. Studies of differences in gene expression were carried out with 24 K oligonucleotide microarrays for genome-wide gene expression analysis and the real-time RT-PCR technique for microarray validation. To determine the physiological processes that are differentially regulated between strains, the DAVID 6.7 Functional Annotation Tool was used.Gapdh and Hprt1 were used as a reference. These genes were not regulated between HA and LA during stress procedure in our study and have been used successfully as a housekeeping control in previous study as well . Large-scale study of real-time RT-PCR and gene expression measurements with commercial oligonucleotide microarrays concluded that microarrays are invaluable discovery tools with acceptable reliability for genome-wide gene expression screening, though validation of putative changes in gene expression remains advisable  and low (LA) swim stress-induced analgesia inbred mouse strains. The strains display robust differences in behavioral tests reflecting depression and in responses to different classes of antidepressants  . Transthyretin is one of the three prealbumins: alpha-1-antitrypsin, transthyretin, and orosomucoid. Transthyretin is a carrier protein and a major transporter of thyroid hormones and retinol in the plasma and cerebrospinal fluid  in both high and low analgesia mouse strains.The chronic mild stress has been studied at the cellular level  in HA mice and increased protein kinase C, gamma (Prkcg) in LA. Glucocorticoid release during stress could potentiate stress-signaling pathways through activation of PKC signaling . Moreover, PKCs require Ca2+, diacylglycerol (DAG), and a phospholipid such as phosphatidylserine for activation. According to this, another interesting group of genes identified in LA mice was intracellular signaling cascade/GTP-ase regulator activity cluster. This cluster, besides Prkcg consist of several upregulated genes in LA CMS animals including diacylglycerol kinases epsilon (Dgke) and gamma (Dgkg) involved in PKC signaling.Chronic stress alters catecholamine pathways increasing noradrenergic innervation of the PFC although dopamine becomes depleted with severe chronic stress  and Cntnap4 (contactin associated protein-like 4). Crmp1 encodes a member of a family of cytosolic phosphoproteins expressed exclusively in the nervous system. Encoded protein is thought to be a part of the semaphorin signal transduction pathway implicated in semaphorin-induced growth cone collapse. CRMP1 is necessary for signaling by class 3 semaphorins and subsequent remodeling of the cytoskeleton. Furthermore, it plays a role in axon guidance while Cntnap4 product belongs to the neurexin family, members of which function in the nervous system as cell adhesion molecules and receptors  in HA CMS mice and downregulation of metabotropic glutamate receptor 5 (Grm5) in LA CMS mice. Grm5 is a subtype of group I glutamate receptors, is highly expressed in limbic forebrain regions, and is thought to modulate anxiety-related processes. The blockade of Grm5 by specific antagonist 2-methyl-6-(phenylethynyl) pyridine, reduce extracellular norepinephrine, the impact of which may contribute to their anxiolytic actions . Activation of the hypothalamic-pituitary-adrenal system by psychosocial stress is accompanied by an increase in peripheral plasma AVP levels in human study  in hippocampus as compared to naïve LA mice  in HA mice after CMS procedure. This gene encodes a common precursor for two peptides, neuromedin N and neurotensin. Neurotensin is a secreted tridecapeptide, which is widely distributed throughout the central nervous system, and may function as a neurotransmitter or a neuromodulator. It may be involved in dopamine-associated pathophysiological events. Both, Nts and Drd1a were clustered in group of genes involved in homeostatic process/regulation of biological quality in HA CMS mice. In addition to the role of NT in the regulation of nociceptive processing, there is accumulating evidence suggesting that NT is involved in hormonal, neural, and behavioral stress-related responses. NT turnover is also altered under physiological stress. Exposure to various stressors, such as immobilization or cold-water swim, induces an up-regulation of the NT precursor mRNA in several hypothalamic regions, including the medial preoptic area and the paraventricular nucleus  receptor transcript  and downregulation of corticotrophin-releasing hormone receptor 1 (Crhr1). Calpains are a group of calcium-dependent protease that plays a significant role in synaptic plasticity, cell motility, and neurodegeneration , CRH receptors , mineralocorticoid, and glucocorticoid receptors  (Joels and Baram Prlr). Study showed that prolactin (PRL) is a neuromodulator of behavioral and neuroendocrine stress coping in the rat ESM 2(XLS 55 kb)ESM 3(XLS 61 kb)Below is the link to the electronic supplementary material.


Cardiovascular magnetic resonance (CMR) using T2-weighted sequences can visualize myocardial edema. When compared to previous protocols, newer pulse sequences with substantially improved image quality have increased its clinical utility. The assessment of myocardial edema provides useful incremental diagnostic and prognostic information in a variety of clinical settings associated with acute myocardial injury. In patients with acute chest pain, T2-weighted CMR is able to identify acute or recent myocardial ischemic injury and has been employed to distinguish acute coronary syndrome (ACS) from non-ACS as well as acute from chronic myocardial infarction.T2-weighted CMR can also be used to determine the area at risk in reperfused and non-reperfused infarction. When combined with contrast-enhanced imaging, the salvaged area and thus the success of early coronary revascularization can be quantified. Strong evidence for the prognostic value of myocardial salvage has enabled its use as a primary endpoint in clinical trials. The present article reviews the current evidence and clinical applications for T2-weighted CMR in acute cardiac disease and gives an outlook on future developments."The principle of all things is water"Thales of Miletus (624 BC - 546 BC) Cardiovascular magnetic resonance (CMR) is well-established and increasingly used in clinical practice for the diagnosis and management of cardiovascular disease -3.Importantly, recent technological advances of CMR have introduced its use for visualizing certain tissue changes in patients with acute myocardial diseases. This is of particular interest in patients with suspected ischemic disease, a broad and heterogeneous population that challenges the clinician in terms of: 1) accurately establishing the diagnosis; 2) risk stratification; 3) therapeutic decision making; and 4) monitoring response to therapy . On. On46]. Another limitation is the often qualitative nature of T2-weighted imaging. Interpretation depends on regional differences in myocardial signal intensity, which may vary depending on sequence parameters . Especially when comparing edema with scars, cut-off values for defining abnormal vs. normal tissue for automated quantification are not sufficiently standardized between methods.As mentioned above, two recently introduced techniques address several of these limitations, but these new methods are also qualitative and still depend on subjective interpretation of T2-weighted images and relative regional differences in myocardial signal intensity. Particularly, direct measurement of T2 by mapping techniques could overcome such limitations . FurtherIn summary, CMR protocols for edema detection still require further optimization. Despite these challenges, however, T2-weighted CMR usually provides images with diagnostic quality, allowing a clinically useful interpretation in most patients.CMR imaging is emerging as a diagnostic tool for the detection, differential diagnosis, management and prognostication of patients with suspected or established ACS ,19,60,61A recent prospective study demonstrated that T2-weighted CMR improves diagnostic accuracy when characterizing patients with possible ACS who presented to the emergency department with chest pain. Adding T2-weighted imaging to a standard CMR protocol  increased the specificity, positive predictive value, and overall accuracy for detection of an ACS from 84% to 96%, 55% to 85%, and 84% to 93%, respectively [T2-weighted imaging provides incremental diagnostic information above and beyond viability assessment, with acuity of cardiac disease being one of the most important determinants for patient management. Specifically, T2-weighted images can determine the acuity of many patients with unstable angina as early as 30 min after the onset of ischemia, a feature that cannot be ascertained by current generation biomarkers or CMR scar imaging using LGE .in vivo in patients with NSTE-ACS can be a powerful tool with major clinical implications. However, further randomized studies are warranted to study the impact of T2-weighted CMR imaging on selection of management strategies and ultimately on prognosis in patients with NSTE-ACS.A recent study extended this work and demonstrated that CMR with edema imaging has great potential to stratify patients admitted with Non-ST-Segment Elevation (NSTE) ACS by identifying higher-risk patients who would qualify for early invasive management strategies . These fEdema imaging is also useful in patients with troponin-positive chest-pain and unobstructed coronary arteries. In this diagnostic challenging and important group of patients, T2-weighted imaging is complementary to LGE imaging for identifying the underlying etiology. The differential diagnosis includes ischemic infarction, myocarditis, stress-induced cardiomyopathy (Takotsubo) and/or other cardiomyopathies (e.g. dilated cardiomyopathy) ,19,63. Iin vivo T2-weighted images obtained 2 days following left anterior descending coronary occlusion accurately define the ischemic area at risk in both reperfused [Experimental studies  have demperfused  and nonrperfused . The sizFriedrich et al.  applied Furthermore, trials demonstrated that myocardial salvage assessment by CMR is a reproducible tool  that ideWhile myocardial salvage assessed by CMR is independently associated with adverse LV remodelling  and moreTheoretically, there are advantages of measuring the salvaged area at risk over infarct size as an indicator of therapeutic efficacy in clinical trials. It is important to keep in mind that the size of the area at risk and the infarction depend on the size of the perfusion bed. Consequently, these parameters are subject to significant baseline differences between patients and even individual clinical events, independent of other clinical circumstances including revascularization. In clinical trials, this bias may account for more than 50% of infarct size variability ,73. As aRecent studies have also demonstrated the potential of T2-weighted imaging to detect intramyocardial hemorrhage (IMH), a marker of severe reperfusion injury -79. As IRecently also other CMR methods for area at risk measurement have been suggested. One of these methods is the endocardial extent of infarction as assessed by LGE CMR . The patAnother potential CMR method for area at risk measurement is to assess the dysfunctional myocardium in cine SSFP imaging . MyocardLimiting the utility of a functional definition of the area at risk, there is evidence that areas of functional impairment have been found to be distinctly smaller than the area at risk as derived from T2-weighted imaging ,13,65; tTaken together other CMR methods for myocardium at risk assessment have significant limitations. Therefore T2-weighted CMR imaging currently seems to be the most accurate and best validated CMR technique for area at risk measurement.Although T2-weighted CMR technology has improved substantially in recent years, it will further benefit from more robust protocols. T2 quantification (mapping) offers the potential for improved detection of myocardial edema, but further studies are warranted to evaluate the clinical applicability of this technique in the range of conditions that are known to globally or regionally affect the myocardial water status. Furthermore, scanning protocols have to be simplified and consolidated, and post-CMR processing and evaluation procedures need to be standardized and less time-consuming .Future studies should address how much of a myocardial injury is needed to become apparent on CMR images of myocardial edema and how clinical and biological factors (e.g. intensity and/or duration of ischemia) affect the magnitude, duration, and time course of myocardial T2 changes. Furthermore, experimental studies are needed to clarify the relative importance of factors capable of influencing T2 signals during and after ischemia .Studies in large, well-defined subsets of patients will be needed to define the additive value of T2-weighted imaging in specific clinical settings. For example it is unknown if cardioprotective strategies specifically targeting edema would help as adjunctive therapies to improve outcomes. Finally, as for all imaging techniques, there is the need for randomized studies comparing CMR-guided treatment decisions including edema imaging versus standard treatment to better understand how CMR and T2-weighted imaging can improve treatment and outcome in patients with ACS.Imaging myocardial edema with CMR in patients with acute cardiac disease provides useful incremental information on the acuity of myocardial injury, be it ischemic or non-ischemic.in vivo tissue marker for acute myocardial injury also significantly improves the clinician`s ability to stratify the risk in patients with acute chest pain syndromes and triage them to appropriate treatment.In institutions with rapid access to CMR, the assessment of myocardial edema as an Comprehensive CMR scans also allows for obtaining accurate information on LV function, size, morphology, perfusion, and scarring during the same scan , so has to be considered very efficient. Importantly, in combination with scar imaging, T2-weighted CMR of myocardial edema differentiates reversible from irreversible injury and can quantify myocardial salvage after coronary revascularization, with important implications for patient management and prognosis. Furthermore, it provides a very powerful, safe and cost-efficient endpoint for clinical trials on myocardial revascularization.In conclusion, T2-weighted CMR is a validated, unique technique which should be considered as an essential diagnostic tool as part of a comprehensive CMR scan.The authors declare that they have no competing interests.Each author participated in researching the relevant articles and drafting and revising the manuscript.


Exchange between the nucleus and the cytoplasm occurs through nuclear pore complexes (NPCs) embedded in the double membrane of the nuclear envelope. NPC permeability barrier restricts the entry of inert molecules larger than 5 nm in diameter but allows facilitated entry of selected cargos, whose size can reach up to 39 nm. The translocation of large molecules is facilitated by nuclear transport receptors (NTRs) that have affinity to proteins of NPC permeability barrier. Viruses that enter the nucleus replicate evolved strategies to overcome this barrier. In this review, we will discuss the functional principles of NPC barrier and nuclear transport machinery, as well as the various strategies viruses use to cross the selective barrier of NPCs.  The double membrane of the nuclear envelope surrounds the nucleus and separates the cellular genome from the cytosol during interphase. This subdivision leads to the physical separation of transcription and translation, which require a highly coordinated exchange between the cellular compartments. Viruses that replicate in the nucleus also have to pass the nuclear envelope barrier during the infection. Almost all exchange between nucleus and cytoplasm occurs through the nuclear pore complexes (NPCs) that are embedded in the double membrane of the nuclear envelope ,2,3,4,5.NPCs are very large macromolecular assemblies with an approximate mass of 125 MDa in higher eukaryotes ,7. This 3 facilitated translocation events per second [The NPC functions as a highly selective gate and allows passage of molecules in two modes: passive diffusion and facilitated translocation . Passr second . The majority of facilitated translocations are mediated by NTRs of the importin-β superfamily . TheNucleocytoplasmic transport through the NPCs occurs in a step-wise manner: (i) binding of a cargo molecule to its cognate NTR; (ii) docking of NTR·cargo complex to the NPC; (iii) translocation through the nuclear pore; and (iv) cargo release on the opposite side of the nuclear envelope. The directionality of Imp/Exp-mediated transport processes is determined by the RanGTP gradient across the nuclear envelope ,28. Ran’In the cytoplasm, RanGTPase-activating protein RanGAP triggers hydrolysis of Ran-bound GTP ,35. HoweNPCs are composed of approximately 500 individual polypeptides representing multiple copies of about 30 different nuclear pore proteins  . AccordiThe natively unfolded nucleoporin domains are enriched with Phe-Gly (FG), Phe-x-Phe-Gly  or Gly-Leu-Phe-Gly (GLFG) clusters separated from each other by hydrophilic spacer regions of different length. Approximately 1/3 of all nuclear pore proteins are FG nucleoporins . FG nuclN-acetylglycosamine (O-GlcNAc) to Ser or Thr amino acids [A remarkable feature of vertebrate FG nucleoporins is that they are post-translationally modified by the attachment of monomeric residues of O-linked no acids ,56,57,58no acids . This soThe permeability barrier of the pore plays a dual role: it restricts the diffusion of inert molecules, whose mass exceed 20–40 kDa, but allows the passage of large NTR·cargo complexes, whose mass can reach several megadaltons. The facilitated translocation of NTR·cargo complexes through the NPCs barrier is linked to the interactions between NTRs and FG nucleoporins. Several models have been proposed to explain the molecular mechanism of NPC permeability barrier function. The “Brownian/virtual gate” model ,61 propoThe “reduction of dimensionality” model  assumes Xenopus egg extract and recombinant FG Nups demonstrated a requirement for multivalent cohesion between FG repeats to maintain integrity and selectivity of NPC barrier [The “selective phase” model was proposed to explain the molecular mechanism of NPC permeability barrier function. The model assumes that the interactions between the hydrophobic FG-clusters crosslink the FG-repeat domains into a sieve-like FG-hydrogel ,45. Acco barrier . This co barrier . Experim barrier . In addi barrier . Experim barrier . The conViruses that replicate in the nucleus have evolved strategies to go across the NPC. The variety of strategies developed by viruses to enter the nucleus is remarkable, and likely reflects the need to complete earlier steps such as entry, trafficking and uncoating in an orderly fashion before engagement with NPCs. For example, viruses with large genomes are unlikely to shed their capsid shell early post-infection in the cytoplasm, because the intracellular viral complex would become too bulky and too loose for cytoskeletal transport. Indeed herpesviruses and adenoviruses appear to dock their intact capsid shell at the NPC where partial uncoating occurs, and the viral DNA genome is made visible to the nuclear import machinery . HerpesvAnother relevant constraint determining the mechanism of viral nuclear import is the size of the viral capsid shell, which often exceeds the functional diameter of the NPC. Studies employing gold nanoparticles determined that 25 nm was the maximal functional diameter of the NPC . HoweverIn light of these observations, an important question is how large viral capsids can rearrange FG Nups inside the central NPC channel to negotiate the barrier and at the same time maintain the barrier selectively permeable. One possible model predicts that FG Nups exist in two bi-stable conformations: either clustered towards the centre or towards the NPC wall, depending on the strength of the intermolecular interactions  . Large vin situ to expose the nucleic acids [The capsid of some viruses are able to go across NPC more or less intact, suggesting that uncoating can happen in both cytoplasm and nucleus. Indeed there is evidence that adeno-associated virus (AAV), hepatitis B virus (HBV) and Baculoviruses uncoat after nuclear entry or at the nuclear side of NPCs ,80,81, aic acids ,86. The ic acids ,88,89,90Chiromonus salivary glands [Viruses that uncoat their capsid shell in the cytoplasm or at the cytoplasmic side of the NPC must have evolved ways to translocate their nucleic acids across the NPC barrier. This is particularly impressive for the large >150 Kb DNA genomes of herpesviruses. Nucleic acids are negatively charged and hydrophylic, which makes them very unsuited to cross the hydrophobic barrier in the NPC central channel. Thus it is not clear how large viral genomes can cross the NPC barrier but it is possible that at least some mechanisms are conserved with export of mRNA complexes (reviewed in ). Messeny glands , presumay glands . Moreovey glands . CurrentViruses have evolved to exploit a wide range of host factors for nuclear import, although some pathways appear at least partially conserved and in some cases redundant. For example HIV-1 utilizes Nup358, Nup153, Nup98, in addition to NTRs such as Imp7, Impβ and Impα ,100,101.Viruses, many aspects of viral nucleocytoplasmic transport will be discussed in greater depth. We hope this Special Issue will provide a useful and enjoyable source of information and stimulate further research in the field.Viruses exploit or alter nuclear transport processes using a remarkable variety of mechanisms, and therefore they have been intensively studied to clarify the biology of nuclear transport ,109,110.


Histamine intolerance is mainly caused by an imbalance of histamine intake and the capacity for histamine metabolism and degradation. The main enzyme for metabolism of ingested histamine is diamine oxidase (DAO). Determination of DAO activity in serum might be useful for differential diagnosis of histamine intolerance. Over the 3.5-year-long period we have recruited 316 patients with suspicion of histamine intolerance and excluded food allergy together with 20 healthy controls. Serum DAO activity was measured with Enzyme immunoassay for the quantitative determination of histamine-degradation activity by DAO in serum. Twenty patients with histamine intolerance and highly reduced initial activity of serum DAO (<40 HDU/ml) went to a histamine-free diet and after 6 to 12 months of histamine-free diet all clinical parameters and serum for determination of DAO activity were taken again.We found that DAO activity was significantly lower in patients than in healthy control subjects (P<0.0001). Furthermore, 54 patients had highly reduced activity of DAO (<40 HDU/ml). The main symptoms involved the skin, gastrointestinal tract, respiratory system and eyes. In all 20 patients after the histamine-free diet the main clinical symptoms typical for histamine intolerance have disappeared. Furthermore, the measured values for activity of serum DAO have increased significantly (p<0.0001).We can conclude that determination of DAO activity in serum is a useful diagnostic tool, together with detailed history to differentiate between food allergy and histamine intolerance. It should be performed in suspected patients with symptoms like headache, tachycardia, urticaria, pruritus, diarrhea and hypotension, where food allergy was excluded. Furthermore, our results showed the benefit of histamine-free diet, since after the diet majority of histamine related symptoms have disappeared as well as the DAO activity in serum has increased.


Health outcomes are key to patients and clinicians, but trials of alcohol brief intervention (BI) tend to focus on behavioral outcomes, while those finding positive effects on physical- or mental-health outcomes generally lack power to show robust effects. To test the feasibility of a randomized controlled trial of BI for either hypertensive or depressed primary-care patients who drink alcohol above recommended levels, records at 25 English primary-care practices were searched for adult patients with risky drinking and comorbid hypertension or depression. Eligible patients were randomized to either a hypertension or depression arm, then to a control or intervention condition. They were screened for at-risk drinking using the Alcohol Use Disorders Identification Test (AUDIT). Consenting respondents scoring >7 on the AUDIT completed the Patient Health Questionnaire-9 (PHQ-9) (depression arm) or blood pressure measurement (hypertension arm) and received either BI or standard advice. At six-month follow-up, participants are again screened for alcohol use and comorbid condition. Seventeen practices  searched their databases. Fourteen percent of adult patients  drank above guidelines. Twenty percent of adult patients  were hypertensive, of which 5%  also drank heavily. Fourteen percent of adult patients had mild to moderate depression , of which 2%  also drank heavily. Of 2590 eligible patients, 633 (24%) completed the AUDIT. Thirty-five percent scored positively on the AUDIT in the hypertension arm, and 50% scored positively in the depression arm. Eighty patients were recruited to the hypertension arm, and recruitment to the depression arm is ongoing. Patients eligible for an RCT of BI for comorbid heavy drinking and hypertension or mild/moderate depression are identifiable in primary care records, though with variation among practices. Almost a quarter of these patients can be screened by mail for current alcohol use; more screen positively in the depression arm than in the hypertension arm. A trial in the hypertension arm seems most feasible.


Introduction. Pityriasis lichenoides is a rare, acquired spectrum of skin conditions of an unknown etiology. Case Report. A 28-year-old man presented with recurrent outbreaks of herpes simplex virus associated with the onset of red-to-brown maculopapules located predominantly in trunk in each recurrence. Positive serologies to herpes simplex virus type 2 were detected. Histopathological examination of one of the lesions was consistent with a diagnosis of pityriasis lichenoides chronica. Discussion. Pityriasis lichenoides is a rare cutaneous entity of an unknown cause which includes different clinical presentations. A number of infectious agents have been implicated based on the clustering of multiple outbreaks and elevated serum titers to specific pathogens . In our patient, resolution of cutaneous lesions coincided with the administration of antiviral drugs and clinical improvement in each genital herpes recurrence. In conclusion, we report a case in which cutaneous lesions of pityriasis lichenoides chronica and a herpes simplex virus-type 2-mediated disease have evolved concomitantly. Pityriasis lichenoides is a rare, acquired spectrum of skin conditions of an unknown etiology, although infectious agents have been suspected as etiologic factors.A 28-year-old man presented with recurrent outbreaks of herpes simplex virus (HSV) type 2 in the form of ulcers on the glans penis associated with the onset of red-to-brown maculopapules located predominantly in trunk in each recurrence. The patient had four recurrences on a two-year followup .Positive serologies to HSV type 2 were detected in each outbreak (IgM and IgG). He received acyclovir 400 mg five times a day for 5 days, then a prophylactic dose of 400 mg twice a day; however, after discontinuation of antiviral drug, lesions reappeared.Histopathological examination of one of the lesions showed epidermal acanthosis, scanty lymphocytic exocytosis, without spongiosis, or focal necrosis. Perivascular infiltrates of lymphocytes with occasional eosinophils and erythrocytic extravasation were seen in the dermis . HSV DNAPityriasis lichenoides (PL) is a rare cutaneous entity of an unknown cause which includes different clinical presentations. It encompasses a spectrum of clinical presentations ranging from acute papular lesions rapidly evolving to form pseudovesicles and central necrosis (pityriasis lichenoides et varioliformis acuta or PLEVA) to small, scaling, benign-appearing papules (pityriasis lichenoides chronica or PLC). PLEVA and PLC are not distinct diseases but are rather different manifestations of the same process. The pathogenic mechanism behind PL is unclear although infectious or drug-related hypersensitivity reactions versus lymphoproliferative disorders are the mainstay theories. A number of infectious agents have been implicated based on the clustering of multiple outbreaks and elevated serum titers to specific pathogens, such as human immunodeficiency virus, hepatitis C virus, cytomegalovirus, Epstein-Barr virus, Toxoplasma gondii, and HSV –4, but tIn our patient, resolution of cutaneous lesions coincided with the administration of antiviral drugs  and clinical improvement in each genital herpes recurrence. PLC did not become less severe at successive attacks. Whether this association represents HSV reactivation caused by immune dysfunction related to the skin disease or HSV acting as an antigenic trigger is uncertain.In conclusion, we report a case in which cutaneous lesions of pityriasis lichenoides chronica and a herpes simplex virus type 2-mediated disease have evolved concomitantly.


High-grade astrocytoma remains a significant challenge to the clinician and researcher alike. Intense study of the molecular pathogenesis of these tumors has allowed identification of frequent genetic alterations and critical core pathways in this disease. The use of novel mouse genetic tools to study the consequence of specific mutations in brain has led to the development of multiple representative genetically engineered mouse models that provided novel insights into gliomagenesis. As we learn more about the biology of high-grade astrocytoma from the study of these models, we anticipate that our improved understanding will eventually lead to greater success in clinical trials and improved outcome for patients. High-Grade Astrocytomas (HGAs) are among the most aggressive and deadly malignancies in man. Unlike most cancers, these primary central nervous system (CNS) tumors do not achieve their lethality by metastatic spread to secondary organs, leading to overwhelming tumor burden and multiorgan failure in patients. The devastating effects of HGAs can be traced to multiple contributing factors. 1) The brain is encased within a solid cranial vault such that progressive tumor expansion leads not only to specific neurological signs and symptoms but also to generalized CNS depression and dysfunction. 2) Tumors are often localized within or adjacent to critical brain structures making complete surgical resection neurologically devastating and therefore impossible. 3) HGAs are highly invasive such that even in cases where complete radiographic resection is achieved local and/or distant microscopic spread of tumor cells at the time of diagnosis usually leads to rapid tumor regrowth. 4) Tumor cells and in particular glioma stem-like cells exhibit a high degree of radio and chemoresistance due to both intrinsic  and extrinsic (eg. blood-brain barrier) mechanisms. As a consequence, despite contemporary multimodal therapy that includes aggressive surgery, maximally delivered radiotherapy and temozolomide, median overall survival for glioblastoma, the most advanced form of HGA, is 15-18 months, a dismal prognosis which has not significantly improved in several decades . Th. Th17]. f tumors , 15. NF1ositol 3,,5-trisphf tumors . This suTP53 is the most commonly mutated tumor suppressor gene in human cancer. The P53 pathway regulates the critical checkpoint that detects oncogenic stress and DNA damage, which if unresolved leads to cellular senescence or apoptosis  are targeted in at least 87% of cases [s Figure . In HGA,of cases . Therape1/S boundary  and strikingly, by the absence of TP53 mutations. The mutual exclusivity of HGAs with PDGFR-α expression, EGFR expression and loss of NF1 was also seen in a targeted proteomic study [Whole transcriptome analysis has emerged as an important method to compare tumor samples and may lead to improved patient stratification and determination of prognosis. This method was used to define expression signatures that distinguish HGA patients according to differential survival outcomes in two independent studies , 28. In  Network . Unsuper Network . Integra Network  showed tsubgroup . In contic study .Ultimately, the goals of genome-wide and integrative genomic analyses are to provide a template for accurate molecular classification of HGA that will be highly predictive for prognosis and can be used to stratify patients for optimal therapy.TP53 [CDKN2A [PTEN and in particular amplifications of the EGFR gene are rare in pediatric HGG [PDGFRA (14%) and homozygous deletion of CDKN2A (16%). IDH1 hotspot mutations are not common in pediatric HGG [While the same WHO criteria apply to the classification of pediatric high-grade gliomas (HGGs), several differences in the clinical behavior of patients hint at fundamental differences that may exist in tumor biology. 1) Malignant transformation of low-grade astrocytomas in children has been described but appears to occur at a much lower frequency than observed in adult patients . 2) A siTP53  and CDKN [CDKN2A  occur witric HGG . These otric HGG -40. The tric HGG , 41. OvePDGFRA amplifications which clustered in the Proliferative subgroup  there was little correlation between focal genetic aberrations and specific subgroups. Furthermore, copy number alterations targeting genes within core pathways were clearly present in pediatric HGG at lower frequency than in adult tumors. This suggests that these tumors utilize alternative mechanisms to dysregulate the pathways or that core pathways play a diminished role in pediatric HGG. Collectively, these studies show similar gene expression-defined subgroups in pediatric and adult HGGs that are driven by a different set of low-frequency genetic events in children.Unsupervised gene expression analysis identified three subgroups corresponding to the Proneural, Proliferative and Mesenchymal subclasses seen in adult HGA . HoweverGFAP-CreER conditional mouse line that drives loxP-dependent recombination in astrocytes and adult neural precursors, cooperativity between the three tumor suppressor genes, Trp53, Pten and Rb1, for gliomagenesis was explored in the context of the adult brain [Pten alone or in combination with Rb1 was unable to initiate glioma formation. In contrast, Trp53 deletion in conjunction with loss of the Pten or Rb1 tumor suppressor gene resulted in high frequency HGA, while deletion of Trp53 alone resulted in HGA with low frequency and with prolonged latency. The importance of concurrent dysregulation of the three core pathways was confirmed as HGAs resulting from targeted deletion of Trp53 and Pten acquired secondary mutations in Rb pathway genes while Pi3k/Akt pathway activation was seen in tumors arising from Rb1 and Trp53 conditional ablation. Accordingly, mice engineered with targeted deletion of all three tumor suppressor genes developed HGA with a significantly reduced latency.As our understanding of genetic abnormalities underlying HGAs becomes more comprehensive, increasingly complex GEMMs have been developed to investigate the contribution and interplay between driver mutations Table . Severallt brain . This stTrp53 and Pten co-deletion resulted in the development of HGA in mouse brains whereas combined deletion of Trp53 and Rb1 or all three together generated primitive neuro-ectodermal tumors (PNETs). Interestingly, overexpression of Cdk4 was present in the Trp53; Pten deleted HGAs demonstrating that Rb-mediated regulation of cell-cycle progression is a core pathway in these tumors. The selective advantage for the development of PNET in this setting likely reflects a less specific manner in which cells are targeted for Cre-mediated recombination. Indeed, accumulating experimental evidence indicates that different types of brain tumors, and even different subgroups within a histological diagnosis, arise as a consequence of specific driver mutations occurring in specific susceptible cell populations [Trp53, Rb1 or Pten, the cell being targeted as well as the developmental context in which mutations occur, are likely to be critical determinants of the resulting tumor type.Using a different approach to target tumor suppressor genes for deletion in post-natal neural progenitor cells resulted in divergent results. Cre-expressing adenovirus was directly injected into the lateral ventricle of mice carrying various combinations of floxed tumor suppressor gene alleles . Only thulations , 45. It Trp53 in embryonic neural stem cells was mediated by a GFAP-Cre transgene [The importance of developmental context is apparent in a study in which an intragenic deletion of ransgene . HGAs wePten deletion alone does not lead to gliomagenesis, activation of other pathway intermediates on their own is similarly inefficient at initiating HGAs. Neither virally-induced expression of the activated EGFR variant, EGFRvIII [Cdkn2a and deletion of Pten in the striatum, EGFRvIII expression resulted in robust tumor formation [Cdkn2a or Trp53 coupled with expression of PDGF-B efficiently initiated HGAs in adult mice [Cdkn2a gene but not by Trp53 deletion.The contributions of various intermediates of the RTK/Pi3k/Akt pathway have been explored in GEMMs. Consistent with the finding that EGFRvIII , nor PDGEGFRvIII  was sufformation . Likewisult mice . Underscult mice . The proNf1 tumor suppressor gene in gliomagenesis has also been investigated using GEMMs. Conditional ablation of the gene in the developing brain using a GFAP-Cre driver mouse line was insufficient to initiate gliomagenesis [Trp53, malignant gliomas were efficiently produced [Nf1 and Trp53 with or without Pten loss in adult neural progenitors gave rise to HGAs [The role of the agenesis  however,produced , 52. As produced . These iRB1 mutations [TP53, but not PTEN or RB1, are found in low-grade astrocytomas prior to malignant transformation [TP53 would appear to be an early mutation in both primary and secondary glioblastoma.Collectively, these results support the notion that three core pathways play critical roles in human and mouse HGA. The experimental evidence that loss of p53 function, but not Pten or Rb, contributes to early stages of gliomagensis in mice is consistent with the elevated incidence of astrocytomas in individuals with Li-Fraumeni syndrome but neither in Pten Hamartoma Tumor Syndrome (PHTS) nor in patients with germline utations -56. Furtormation . TherefoTrp53 and Pten or Trp53, Pten and Rb1 were subjected to genome-wide molecular profiling [Met, Pdgfra and Egfr and genes encoding the upstream Rb regulators Cdk4, Cdk6 and Ccnd1. Meanwhile, gene expression analyses segregated tumors into three subgroups with significant identity to the Proneural, Proliferative and Mesenchymal subgroups defined by human glioblastoma gene expression studies [All of the GEMMs described reproduce human histological features characteristic of aggressive HGAs. However, molecularly targeted therapies are now the focus of many clinical HGA trials with the expectation that they will eventually augment or replace our current treatment regimens. It is therefore critically important for preclinical models of the disease to mirror not only the histology of human astrocytomas but also their molecular pathology. To address this, tumors arising from adult mouse brains with targeted deletions of rofiling . These a studies , 29.Trp53 and Pten as well as in Trp53, Pten and Rb1 mutated tumors. Importantly, these amplifications appeared to drive maximal signaling through the Pi3k pathway suggesting that ablation of Pten function, the principal negative regulator of Pi3k signaling, is not sufficient to activate the pathway. HGAs with concurrent mutations of PTEN and RTK genes have been noted in several studies [It is particularly interesting to note that in these murine astrocytomas, multiple mutations within core pathways were frequently observed . RTK gen studies , 58. Fur studies , a pheno studies , 59, 60. studies  and huma studies , 59. ThePten or the combination of Pten and Trp53 were conditionally targeted. Gene expression analyses revealed that all tumors expressed a signature strongly associated with the Proneural subgroup which is consistent with PDGFRA amplified tumors in adults [A recent study employed a retrovirus encoding PDGF-B and the Cre recombinase which was directly injected into the subcortical white matter of adult mice . HGAs arn adults , 31. Then adults .These studies demonstrate that GEMM HGAs can recapitulate not only tumor histology accurately but also their molecular pathology. Depending on their design, the models can replicate a spectrum of human HGA subtypes or phenocopy specific ones , 61. As et al. suggest that HGAs in their model originate from the Gfap-positive neural precursor cell (NPC) in the subventricular zone and/or from Olig2-expressing progenitor-like cells most commonly found in the corpus callosum [GEMMs offer a unique opportunity to analyze the early events of gliomagenesis which cannot be accurately determined in studies of human tumors or with xenograft models. Careful analysis of early time points in these mouse models coupled with clever tools to trace transformed cells has resulted in a refined understanding of the cell-of-origin of astrocytomas. By analyzing cells that accumulate mutant p53 soon after Cre-mediated recombination, Wang callosum .GFAP-Cre driver line in conjunction with an elegant genetic tag to label cells that had undergone homozygous deletion of Trp53 and Nf1 [However, a recent report utilized the same  and Nf1 . These i and Nf1 . Interes and Nf1 , 47, 48.GFAP-Cre that have activity in the vast pool of embryonic neural stem cells allow for the study of early events of tumorigenesis in these cells and their derivatives. Due to this embryonic activity, there is a strong bias in these models for transformation of the embryonic neural stem cell population over more mature cell types. The GFAP-CreER driver line allows for widespread and unbiased genetic manipulation in mature astrocytes and progenitor cells from all regions of the central nervous system and has revealed tumor formation in non-proliferative zones such as the cortex, brain stem, cerebellum and spinal cord. It is important to note that human HGAs arise predominantly in these non-proliferative regions of the brain. However, identifying the cell-of-origin in these models is challenging and will require additional genetic markers. Further study of these different GEMMs will continue to contribute to our understanding of HGA initiation and progression.These results highlight the differences in experimental approach between the various GEMMs and what we can learn from their use Table . TargeteHGA continues to be one of the most challenging malignancies that patients and clinicians face. It is clear that substantial improvements in neurosurgical techniques, perioperative care and delivery of radiotherapy have failed to appreciably improve patient outcome . LikewisPerhaps more importantly, comparative and cross-species analyses of tumors arising from the various models will help elucidate biological details about HGAs that will be informative for patient selection and stratification during these trials Figure . Future