Antiretroviral therapy (ART) currently enables long-term survival of persons living with HIV (PLWH), but the respite from escalating pandemic AIDS-related deaths is undermined by both emergence of drug-resistant HIV and failure to develop HIV vaccines . Such reAlthough binding of host self antigens by antibodies risks host damage, such binding may occur without resulting in appreciable harm. In support of this view, an apparent lack of pathological manifestations has been noted among healthy individuals who developed circulating antiplatelet autoantibodies  subsequent to immunization with recombinant HIV gp160 ; and cerin vivo, to block infection without producing excessive damage. Such targeting of host self epitopes might be sufficient to block infection ; otherwise, infection still might be blocked by simultaneous binding of both host self- and pathogen-associated epitopes by synergistically acting antibodies . Furthermore, host self epitopes tend to be highly conserved, which could facilitate vaccine design for entire host populations. The risk–benefit trade-off posed by host-reactive antibody-mediated immunity could be explored initially through passive immunization with monoclonal antibodies (mAbs), before committing to active-immunization strategies . The mAbs could be developed in tandem with specific antidotes  for treating any adverse reactions that might occur, for instance, due to effector mechanisms such as complement activation and antibody-dependent cell-mediated cytotoxicity (ADCC). Antibody-mediated complement activation might be minimized by avoiding the juxtaposition of target epitopes , whereas damage due to any activated complement could be mitigated by complement-inactivating agents ; and damage due to ADCC might be addressed by suppression of natural killer (NK) cell activity. Such complications could pose barriers to regulatory approval, albeit perhaps less so for therapeutic versus prophylactic vaccines particularly where net benefit  would be strongly compelling. Nevertheless, either or both complement activation and ADCC still might contribute to net benefit if they actually decreased pathogen replication [e.g., possibly with antibodies to the scavenger receptor CD36, which have been shown to inhibit HIV-1 release from infected macrophages by clustering newly formed virions at their site of budding , particularly against human rhinovirus (for which ICAM-1 serves as the major host receptor) . Anti-inThe scheme described herein thus shifts the focus of immunity-oriented approaches in health care, from immune destruction of specific targets back to the original object of vaccination, namely host protection against disease. This widens the scope of vaccines and immunotherapeutics, by placing due emphasis on immune targeting of host self epitopes as a potential means for host protection associated with negligible or justifiably limited host damage. More generally, immune targeting of epitopes may be broadly conceptualized in terms of high-level functional outcomes including both familiar consequences of conventional immunization regimens  as well as less obvious and possibly even counterintuitive but nonetheless beneficial results . Such a perspective provides the basis for an expanded paradigm of biomedically enhanced immune function, the essence of which is concisely expressed as the idea of epitopes for protective immunity targeting antigens of pathogen and/or host (EPITAPH).The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Liposarcoma (LPS) is rare malignant tumor of fat cells in deep soft tissue that affects adults between the ages of 40 and 60 . This tuWDLPS and DDLPS subtypes have a characteristic feature of amplified region of chromosome 12q13-15 containing several well-known oncogenes such as MDM2, CDK4 and HMGA2 . We founNext, whole exome sequencing and targeted exome sequencing identified various known cancer related and novel genes recurrently mutated in different subtypes of LPS. MAPK, ErbB, JAK-STAT, Wnt, apoptosis, cell cycle, DNA replication and repair and axon guidance pathways were found to be potentially involved in liposarcomagenesis. We identified recurrent mutations in previously unidentified genes in LPS associated with DNA damage repair pathways. LPS tumors with DNA repair mutations could be completely dependent on other backup repair pathways for the survival which may be exploited to induce synthetic lethality as therapeutic approach in these tumors.Interestingly, we reported for the first time multi-region genomic analysis of single LPS patient's tumor signifying intra-tumor mutational and copy number heterogeneity. The current basis for most of the personalized medicine approaches depends on the genomic landscape of a single tumor biopsy sample. Due to the frequent large size of LPS tumors compared to other solid tumors, intra-tumor heterogeneity will lead to difficulties in identifying biomarkers and therapeutic targets. Our preliminary analysis of intra-tumor heterogeneity mandates the need for future detailed studies exploring the evolution of LPS tumors leading to progression.In summary, our recent work gave insights into global genomic spectrum of LPS cohort for development of novel therapeutic strategies and for understanding the pathogenesis this deadly disease.


Thymallus arcticus) and rainbow trout (Oncorhynchus mykiss) exhibited kilometer-scale movements among streams during the summer growing season. Although we monitored movements at a small fraction of all tributaries used by grayling and rainbow trout, approximately 50% of individuals moved among two or more streams separated by at least 7 km within a single summer. Movements were concentrated in June and July, and subsided by early August. The decline in movements coincided with spawning by anadromous sockeye salmon, which offer a high-quality resource pulse of food to resident species. Inter-stream movements may represent prospecting behavior as individuals seek out the most profitable foraging opportunities that are patchily distributed across space and time. Our results highlight that large-scale movements may not only be necessary for individuals to fulfill their life-cycle, but also to exploit heterogeneously spaced trophic resources. Therefore, habitat fragmentation and homogenization may have strong, but currently undescribed, ecological effects on the access to critical food resources in stream-dwelling fish populations.Stream-dwelling fishes inhabit river networks where resources are distributed heterogeneously across space and time. Current theory emphasizes that fishes often perform large-scale movements among habitat patches for reproduction and seeking refugia, but assumes that fish are relatively sedentary during growth phases of their life cycle. Using stationary passive integrated transponder (PIT)-tag antennas and snorkel surveys, we assessed the individual and population level movement patterns of two species of fish across a network of tributaries within the Wood River basin in southwestern Alaska where summer foraging opportunities vary substantially among streams, seasons, and years. Across two years, Arctic grayling ( River networks are hierarchically structured systems characterized by a continuum of downstream changes in biota and ecosystem processes, which generate coarse-scaled patterns of biotic and abiotic heterogeneity ,2. HowevMovement among habitats is one strategy animals have evolved to track spatially and temporally variable resources. For freshwater fishes, it has been widely acknowledged that relatively long-distance movements are necessary for the long-term persistence of populations . HoweverThymallus arcticus) and rainbow trout (Oncorhynchus mykiss) are two species of non-anadromous (resident) salmonids that co-occur in streams and rivers throughout western Alaska. Grayling and rainbow trout play a large ecological and economic role in the region, as they can often comprise the majority of resident stream biomass and are the mainstay of recreational fisheries )It is interesting to consider why movements at larger spatial scales are rarely documented in freshwater fishes. One explanation is that the range of observed movement distances will directly depend on the interplay between the finest spatial and temporal resolution studied (grain) and the size of the entire study area or duration  by year and stream. Only "returning" fish were used in movement analyses.(DOCX)Click here for additional data file.S1 Figin (A) Hidden Creek, (B) Lynx Creek, (C) Teal Creek, and (D) Stovall Creek.(TIF)Click here for additional data file.S1 File(DOCX)Click here for additional data file.


Technology in cataract surgery is constantly evolving to meet the goals of both surgeons and patients. Recent major advances in refractive cataract surgery include innovations in preoperative and intraoperative diagnostics, femtosecond laser-assisted cataract surgery (FLACS), and a new generation of intraocular lenses (IOLs). This paper presents the latest technologies in each of these major categories and discusses how these contributions serve to improve cataract surgery outcomes in a safe, effective, and predictable manner. With cataract surgery regarded as the most widely performed surgical procedure, a demand exists for continued innovation and technology. The latest advances evolved through application of well-defined principles to current surgical goals and patient expectations. For example, femtosecond laser technology emerged after fifty years of employing laser technology in ophthalmology . TheodorMore than ever, patients have the desire to reduce their dependence on spectacles after cataract surgery. Physicians now have access to advanced diagnostics that can better quantify conditions such as dry eye, light scatter, and posterior corneal astigmatism. These technologies can enhance refractive measurements and appropriate IOL selection.McDonald recently reported that the postoperative prevalence of dry eye related symptoms is approximately 88% . AnalysiEvaluation of optical quality also aids in decision making between corneal or lens-based procedures. The C-Quant  assesses straylight subjectively by utilizing a compensation comparison method . The AcuBoth anterior and posterior corneal astigmatism should be taken into account in IOL planning, particularly in patients desiring astigmatic correction. Inaccuracies arise when posterior corneal astigmatism is measured based on the assumption of a fixed-ratio relationship with the anterior curvature . The CasPatients with a history of corneal refractive surgery expect reduced dependence on spectacles after cataract surgery. The use of intraoperative aberrometry can adjunctively guide the future of IOL power selection . The OptThe use of the electroretinogram (ERG) has been well described and may have a novel application for refractive cataract surgery. Dr. Richard Mackool described the use of flash ERG testing with office-based electroretinography  in preoperative cataract evaluation. It can provide an objective evaluation of macular function and could be useful in influencing lens selection for patients with conditions such as epiretinal membrane, diabetic retinopathy, and age-related macular degeneration. More studies evaluating ERGs in preoperative cataract assessment need to be done to further assess its value and implications .Femtosecond laser-assisted cataract surgery (FLACS) has realized increasing popularity. Noninferiority has been established relative to manual cataract surgery, and some reports have suggested superiority relative to manual methods . Potenti™ by Alcon), Mayer et al. showed that redocking was unnecessary when a modified soft interface was used, even though some cases resulted in incomplete incisions requiring manual opening [The docking process using the femtosecond laser-eye interface uses a suction ring to stabilize the eye, thereby allowing imaging and laser delivery through a clear optical pathway. Considerations for docking include complete coupling, patient comfort, intraocular pressure elevation, and minimal distortion of anatomy to avoid disruption of the beam path. In a study using Alcon's LenSx platform to compare curved direct contact and modified soft interfaces ; Infinite Vision IOL ; and mechanically adjustable IOLs . These are currently being studied in vivo with promising preliminary results. The second category includes magnetically adjustable IOLs , light adjustable IOLs , and the Perfect Lens . The latter is a novel platform, which can be adjusted with the femtosecond laser based on the concept of refractive index shaping. The light adjustable IOL is currently in FDA clinical trials. Its mechanism involves the use of infrared light to polymerize photosensitive silicone macromers, which results in changes in lens morphology and optical properties .A new generation of optics with extended depth of focus, multifocal rotational symmetry and asymmetry, and accommodating capabilities offers promising strategies to advance functional vision in refractive cataract surgery. The Tecnis Symfony (AMO) is an extended depth of focus IOL that works by correcting chromatic aberration using diffractive optics and reduces glare and halo symptoms classically associated with conventional multifocal IOLs . MultifoAccommodating IOLs aim to simulate the natural mechanism of accommodation. The FluidVision IOL  has silicone oil inside the lens that moves in response to ciliary contraction forces. An electroactive IOL with a liquid crystal that is sensitive to electric current is also in development . Dual accommodating IOLs designed for sulcus placement include the DynaCurve IOL  and the Lumina IOL  . The injThe future of refractive cataract surgery is exciting; in time, these new technologies may be the standard of care. With refinements of the latest technology, FLACS and other parallel advances will provide surgeons with the potential to perform an even safer, predictable, and effective surgery.


Escherichia coli (E. coli) strains that cause ABU have evolved from uropathogenic E. coli (UPEC) by reductive evolution and loss of the ability to express functional virulence factors. For instance, the prototype ABU strain 83972 has a smaller genome than UPEC strains with deletions or point mutations in several virulence genes. To understand the mechanisms of bacterial adaptation and to find out whether the bacteria adapt in a host-specific manner, we compared the complete genome sequences of consecutive reisolates of ABU strain 83972 from different inoculated individuals and compared them with the genome of the parent strain. Reisolates from different hosts exhibited individual patterns of genomic alterations. Non-synonymous SNPs predominantly occurred in coding regions and often affected the amino acid sequence of proteins with global or pleiotropic regulatory function. These gene products are involved in different bacterial stress protection strategies, and metabolic and signaling pathways. Our data indicate that adaptation of E. coli 83972 to prolonged growth in the urinary tract involves responses to specific growth conditions and stresses present in the individual hosts. Accordingly, modulation of gene expression required for survival and growth under stress conditions seems to be most critical for long-term growth of E. coli 83972 in the urinary tract.During asymptomatic bacteriuria (ABU), bacteria colonize the urinary tract for extended periods of time without causing symptoms of urinary tract infection. Previous studies indicate that many  Normal flora furnishes the host with ecological barriers that prevent pathogen attack while maintaining tissue homeostasis. Despite their vast numbers and staggering molecular complexity, microbiomes of the gut, respiratory and urogenital tracts persist without triggering a destructive host response. This lack of destructive inflammation is fascinating and important, as it reflects exquisite molecular regulation of the host environment by commensal bacteria. Simultaneously, the host regulates permitted and unwanted paths of immune activation by discriminating attacking pathogens from beneficial commensals.The failure of asymptomatic carrier strains to trigger disease-associated signaling pathways and pathology has generally been attributed to their lack of virulence and, until recently, it was not clear if, in addition, asymptomatic carrier strains enhance their persistence by actively modifying the host environment. We have discovered that commensal bacteria modulate host gene expression to ensure that destructive immune activation will not occur . These iUTI constitutes a highly relevant model of microbial adaptation, in which the contrasting roles of pathogens and commensals are clearly displayed . PathogeEscherichia coli (UPEC) activate an innate immune response through virulence factor–specific TLR4 signaling, the TRIF/TRAM adaptors, MAPK-, p38- and CREB phosphorylation and IRF3/IRF7, AP1-dependent transcription. We have shown that −/−Irf3 mice develop severe, acute symptoms accompanied by urosepsis and renal abscess formation, demonstrating that the innate immune response orchestrated by IRF-3 is crucial for bacterial clearance and for renal tissue integrity. Human disease relevance is suggested by an increased frequency of functionally relevant IRF3 promoter polymorphisms in about 70% of patients with recurrent acute pyelonephritis . The. The7]. oolgirls  and was E. coli strain VR50, have been also shown to evolve from commensal strains by gaining colonization factors [Bacteria increase their fitness in new host niches by rapid adaptation to changing environmental conditions. They lose or gain genetic material and, through selection, new variants become fixed in the population. Evolution has mainly been assumed to favor virulence, which promotes host-to-host spread and tissue attack. Until recently, evolution of commensalism had not been considered. We have proposed that ABU strains evolve towards commensalism in human hosts, as defined by a reduction in overall genome size, inactivation of virulence genes and modifications of transcriptional regulators ,11. Some factors .E. coli 83972 genome revealed a common ancestry with uropathogenic E. coli strains but a smaller genome size due to multiple deletions and mutations, suggesting that this strain adapted to the human urinary tract by undergoing reductive evolution [fim deletion and several papG point mutations abolish fimbrial expression and adherence [Epidemiologic studies have established that the severity of UTI reflects the virulence profile of the infecting strain, with a higher frequency of tissue-attacking virulence factors expressed by UPEC strains than by most strains causing ABU ,14, despvolution ,16 . Reisolates with slower growth rates in urine were also less competitive relative to ancestor strain 83972. Many reisolates formed less biofilm than parent strain 83972. This phenotype could be correlated with motility and flagella expression [E. coli is modulated in response to the individual host. Further genome-wide, transcriptome and proteome analysis proved that these genomic changes altered bacterial gene expression, resulting in unique adaptation patterns in each patient affecting iron uptake strategies as well as protection against oxidative or nitrosative stress, general stress response, and utilization of different carbon sources [Diverse pheno- and genotypic comparisons uncovered striking differences among patients  . So farE. coli 83972 monoculture populating the bladder [in vivo growth in the urinary tract could include phenotype switching. Alternatively, the occurrence of heterogeneous populations at symptomatic episodes may represent spontaneous stochastic events including minor transient populations [Interestingly, we observed phenotypic variation in the  bladder . This ph bladder  suggests bladder . Small c bladder . Therefoulations .E. coli 83972 colonizes the human urinary tract without inducing a strong immune response and can even actively suppress host gene expression. Genome comparison demonstrated that this strain is a deconstructed, attenuated uropathogen. Although we were able to shed some light on bacterial strategies and conditions which may promote attenuation and E. coli adaptation during prolonged colonization of the urinary tract, specific genomic features of strain 83972, which may account for all aspects of the asymptomatic bladder colonization, have not yet been identified. The detailed analysis of the molecular mechanisms required for the specific bacterium-host interaction resulting in a weak host response will be an important task for future studies to understand how this strain actively modifies the host environment in order to promote persistence.


Identification of disease causing mutations in genetically heterogeneous conditions such as intellectual disability by Sanger sequencing is time-consuming, costly and often unsuccessful. The advent of NGS techniques is paving the way for novel large scale approaches with an unforeseen diagnostic power. However, the plethora of variants of unknown significance detected by genome-wide approaches requires distinctive strategies to identify actually disease-related mutations. We recently showed that exome sequencing of patient-parent trios in sporadic cases of unspecific severe intellectual disability may unravel disease causing mutation in more than 50% of previously unsolved cases. Thereby it became also evident, that the current descriptions of phenotypes associated with mutations in a certain gene, are heavily biased towards certain recognizable patterns. However, while whole exome sequencing may currently provide theoretically the highest cost-efficient diagnostic power, it may miss mutations due to incomplete coverage of certain genes. Therefore in some phenotypes a “clinical exome” limited to a set of genes with currently known monogenic mutations may also be useful.


Prostate cancer remains significant public health concern amid growing controversies regarding prostate specific antigen (PSA) based screening. The utility of PSA has been brought into question, and alternative measures are investigated to remedy the overdetection of indolent disease and safeguard patients from the potential harms resulting from an elevated PSA. Multiparametric MRI of the prostate has shown promise in identifying patients at risk for clinically significant disease but its role within the current diagnostic and treatment paradigm remains in question. The current review focuses on recent applications of MRI in this pathway. An estimated 233,000 newly diagnosed cases of prostate cancer (CaP) are estimated for 2014, with a projected 29,480 CaP deaths in the same year [The shortcomings of PSA and prostate biopsy have propelled the search for alternative measures to improve its diagnostic yield and efficiency. Transrectal ultrasound (TRUS) guided biopsy of the prostate currently remains the gold standard for tissue diagnosis but itself has limitations. The systematic yet random and blinded sampling of TRUS biopsy served a useful purpose from its introduction in the 1980s, when biopsy of palpable, large-volume disease was representative of whole gland pathology. The introduction of PSA and subsequent identification of organ-confined, low volume disease have made TRUS biopsy gradually more antiquated and now CaP remains the only solid organ tumor diagnosed without tumor imaging and a directed sampling method.Multiparametric MRI (mp-MRI) of the Prostate. The role of prostate biopsy has evolved from a purely diagnostic tool to include one that informs clinical decision-making. As such, it is essential to ensure that the appropriate individuals receive a biopsy thus averting unnecessary harm and that biopsy information represents what it truly intends to measure. In an effort to improve on the current standard, PSA-derived markers, PSA kinetics, and patient characteristics including genomic profiling and imaging have all been investigated to refine patient selection with variable results. The application of mp-MRI of the prostate has emerged as a powerful tool that provides detailed anatomical and functional information where it is currently lacking. In addition to high resolution T2 weighted (T2W)  imaging, the integration of diffusion weighted imaging (DWI), dynamic contrast enhanced (DCE) imaging, and spectroscopic  imaging in combination has allowed radiologists to better identify areas of benign and potentially malignant disease  , extracapsular extension (ECE), and even pelvic lymph node involvement. Recent series have reported sensitivity of >80% and specificity of >90 –25. SoylHigh-quality imaging is a fundamental component for patient selection for focal therapy of prostate cancer . Until rThe most valuable utilization of PSA remains after whole-gland treatment, where increases in PSA correlate well with disease recurrence. MP-MRI can be used to identify local recurrence after both radical prostatectomy and external beam radiation with functional imaging sequences driving detection in these patients . The addThe detailed anatomic information afforded from mp-MRI enables its versatile use intraoperatively where PSA falls short. Higher PSA values may provide generalized information, from which inferred risk is calculated based on large datasets but fails to provide patient-specific information that can guide surgery. Improved risk stratification, imaging, and novel therapeutic approaches have enhanced efforts for focal therapy in prostate cancer. Focal therapy techniques including high intensity focused ultrasound and laser induced thermal therapy are largely guided by real-time MRI. MRI can assist in tumor localization and probe placement in addition to treatment monitoring intraoperatively. Novel MR-TRUS fusion platforms for guidance of focal therapy are also beginning to emerge . MR TherOverdetection of clinically insignificant, low-grade, low-volume CaP has spurred the adoption of active surveillance (AS) as an accepted management strategy. AS allows patients with low risk disease to defer definitive therapy, potentially indefinitely, until objective evidence of disease progression is noted. PSA is utilized as both an entry criterion and a mainstay of surveillance in conjunction with random prostate biopsy for AS. MP-MRI and targeted biopsy are able to better differentiate those patients with indolent disease, selecting patients who better represent low-risk disease . Of patiInvestigation of novel technologies harnessing the properties of mp-MRI is showing promise in the staging of prostate cancer. Current models including the UCSF-CAPRA scoring system and the Partin tables integrate PSA to provide valuable insight into presence of metastasis and predictions of organ confined disease. Ultrasmall superparamagnetic particles of iron oxide (USPIO) have shown promise in the preoperative staging of bladder and prostate cancer patients, allowing for detection of metastasis. DWI combined with USPIO has demonstrated potential in discrimination of malignant versus benign pathology in even normal size lymph nodes, though wide adoption has been hindered due to variability in interpretation . In addiFinally, Positron emission tomography (PET) by itself demonstrates high sensitivity for prostate cancer detection but nonspecific uptake in benign lesions limits its diagnostic utility. However, PET/MRI has recently shown promise with a sensitivity and specificity for CaP detection in lesions >5 mm being 84% and 80%, respectively, when correlated with whole mount pathology at radical prostatectomy . FurtherThe novel applications of MP-MRI are encouraging and but are not a panacea to our PSA woes. MRI interpretation is challenging, requiring considerable experience before proficiency. As mp-MRI has gained acceptance, the need for standardized protocols and reporting and high quality studies has been realized . DetermiPSA remains the cornerstone to screening, staging, and surveillance after treatment despite its numerous shortcomings. Currently, mp-MRI is proving an essential adjunct to supplement PSA in proper patient selection for biopsy, treatment, and surveillance. High-quality studies are required to validate the initial encouraging body of literature supporting the use of MRI in this arena.


Information derived from functional magnetic resonance imaging (fMRI) during wakeful rest has been introduced as a candidate diagnostic biomarker in unipolar major depressive disorder (MDD). Multiple reports of resting state fMRI in MDD describe group effects. Such prior knowledge can be adopted to pre-select potentially discriminating features for diagnostic classification models with the aim to improve diagnostic accuracy. Purpose of this analysis was to consolidate spatial information about alterations of spontaneous brain activity in MDD, primarily to serve as feature selection for multivariate pattern analysis techniques (MVPA). Thirty two studies were included in final analyses. Coordinates extracted from the original reports were assigned to two categories based on directionality of findings. Meta-analyses were calculated using the non-additive activation likelihood estimation approach with coordinates organized by subject group to account for non-independent samples. Converging evidence revealed a distributed pattern of brain regions with increased or decreased spontaneous activity in MDD. The most distinct finding was hyperactivity/hyperconnectivity presumably reflecting the interaction of cortical midline structures  with lateral prefrontal areas related to externally-directed cognition. Other areas of hyperactivity/hyperconnectivity include the left lateral parietal cortex, right hippocampus and right cerebellum whereas hypoactivity/hypoconnectivity was observed mainly in the left temporal cortex, the insula, precuneus, superior frontal gyrus, lentiform nucleus and thalamus. Results are made available in two different data formats to be used as spatial hypotheses in future studies, particularly for diagnostic classification by MVPA. Mental disorders featuring depression as a predominant symptom and more specifically major depressive disorder (MDD) are important worldwide public health concerns. In recent years significant progress has been achieved regarding the identification of biological correlates and potential neural mechanisms involved in the pathogenesis of MDD. These scientific efforts comprise studies of genetic foundations, molecular mechanisms including neurotransmitter systems and structural as well as functional neuroimaging  studies in MDD has applied stimulus-based acquisition protocols. Participants were confronted with predefined stimuli in the scanner, e.g., pictures of emotional faces. Brain activity in response to these stimuli was analyzed  or the  amplitude of characteristic low-frequency fluctuations (ALFF or fALFF). Functional connectivity (FC) can be operationalized as the temporal correlation of signal fluctuations in remote brain areas. Conventional FC-analyses are seed-based but FC-analyses in a wider sense include independent component analyses or complex graph theoretical network measures. A minority of studies has applied analyses of effective connectivity   methods have been proposed to identify a subset of most informative features to be used with the aim to increase classification accuracy  and pooled studies with SPECT and PET data  search using the following query on August 20, 2013: (“depression” OR “depressive”) AND  AND .We conducted a PubMed  using the non-additive activation likelihood estimation (ALE) approach with coordinates organized by subject group (ALE-S method) to account for non-independent samples  and 1000 permutations  and a high resolution anatomical template with isotropic voxels in MNI space as distributed with GingerALE.Anatomical labels were automatically assigned in GingerALE. Visualizations were created using Mango  for hypothesis-driven group comparisons and particularly for FS  of spontaneous brain activtiy   or single-photon emission computed tomography (SPECT)  and less consistently the cingulate cortex and thalamus  and derivative techniques: reduced anisotropy measures, a potential marker of fiber integrity, were observed in parts of the superior frontal white matter presumed to connect the dorsolateral prefrontal cortex and anterior cingulate cortex with subcortical nuclei  with lateral frontal areas related to externally-directed cognition. Alterations that can be captured by rs-fMRI seem to differ from those identifiable with other neuroimaging modalities but show considerable overlap. Results of this meta-analysis are provided as coordinates and detailed maps in MNI space to be readily applicable for ROI selection in further rs-fMRI studies in MDD including feature selection for classification approaches with diagnostic intention. By emphasizing spatial precision and sensitivity this approach only provides limited information about the exact functional meaning of altered spontaneous brain activity in MDD.Benedikt Sundermann and Bettina Pfleiderer conceived and designed the study. Mona Olde lütke Beverborg and Benedikt Sundermann identified and screened the articles. Benedikt Sundermann, Bettina Pfleiderer and Mona Olde lütke Beverborg participated in final study selection and group assignment. Benedikt Sundermann and Mona Olde lütke Beverborg conducted the ALE-analyses. Benedikt Sundermann, Bettina Pfleiderer and Mona Olde lütke Beverborg participated in interpretation of the results. Benedikt Sundermann drafted the manuscript. All authors critically revised the manuscript for important intellectual content and approved the final version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Combined PET/MR is a promising tool for simultaneous investigation of soft tissue morphology and function. However, contrary to CT, MR images do not provide information on photon attenuation in tissue. In the currently available systems issue is solved by synthesizing attenuation maps from MR images using segmentation algorithms. This approach has been shown to provide reason-able results in most cases. However, sporadically occurring segmentation errors can cause serious problems. Recently, algorithms for simultaneous estimation of attenuation and tracer distribution (MLAA) have been introduced. So far, validity of MLAA has mainly been demonstrated in simulated data. We have integrated the MLAA algorithm  into thePhantom data were acquired using a whole body phantom with three cylindrical inserts filled with different substances . MLAA-estimated mu-maps of the phantom were compared to the mu-maps resulting from transmission measurements with an ECAT HR+ scanner. We also performed a first qualitative evaluation of the attenuation maps obtained in patient studies.Evaluation of the phantom study showed good concordance between measured and estimated attenuation coefficients for all types of substances used in the phantom. Evaluation of patient data showed some substantial improvements of the MLAA attenuation maps compared to the segmented MR-based attenuation maps.Preliminary results show that for the Philips Ingenuity PET/MR scanner the MLAA algorithm allows to obtain attenuation maps which outperform the MR based maps in several aspects. However, a more detailed analysis is still required to address the question of possible cross-talks in regions with high activity. Additionally, MLAA algorithm substantially increases computational burden leading to long processing times, which makes it currently impractical for clinical application.


Tracheal penetration of esophageal self-expanding metallic stents (SEMS) with/without tracheoesophageal fistula (TEF) formation is a rare occurrence. We report the case of a 66-year-old female patient with advanced esophageal squamous cell carcinoma who had undergone palliative esophageal stenting on three occasions for recurrent esophageal stent obstruction. On evaluation of symptoms of breathing difficulty and aspiration following third esophageal stent placement, tracheal erosion and TEF formation due to the tracheal penetration by esophageal stent were diagnosed. The patient was successfully managed by covered tracheal SEMS placement under flexible bronchoscopy. Esophageal SEMS placement has evolved into a first line approach for the palliative relief of dysphagia for patients with advanced esophageal cancer . The sucDespite the widespread use of esophageal SEMS placement for the numerous previously described indications, delayed prolapse and erosion of esophageal SEMS into the tracheobronchial tree are rarely reported , 3–6. HoA 66-year-old female presented with history of persistent cough and shortness of breath of 2-week duration. Cough used to worsen immediately on intake of liquids/solids and, for the same reason, patient complained of marked reduction in oral intake. There was no history of hemoptysis, wheezing, hoarseness of voice, or chest pain.Patient had been diagnosed with advanced stage carcinoma of the esophagus, 15 months ago. At that time, patient had been evaluated for symptoms of dysphagia, loss of appetite, and significant weight loss. Upper gastrointestinal endoscopy examination (UGIE) performed at that time revealed circumferential growth at 25 cms from incisors and biopsy demonstrated moderately differentiated squamous cell carcinoma. Fine needle aspiration from a palpable right supraclavicular lymph node at that time also confirmed metastatic squamous cell carcinoma. Flexible bronchoscopy (FB) examination at that time was normal. In view of grade III dysphagia and metastatic disease, patient underwent covered esophageal SEMS  placement in November 2012 and palliative chemoradiotherapy , radiotherapy) was concurrently administered. For recurrence of dysphagia six months following esophageal stenting, UGIE was performed which demonstrated tumor in-growth for which a covered SEMS  was placed inside the previous stent. Tumor in-growth recurred nine months later for which another covered esophageal SEMS was placed inside the two previous stents. Symptoms of cough and dyspnea occurred one month following the third esophageal SEMS placement and the patient was referred to the pulmonary outpatient clinic.On examination, patient appeared emaciated. Pulse rate was 100/min, respiratory rate was 25/min, and rest of the vital signs and general physical examination were normal. On examination of the respiratory system, inspiratory crepitations were audible in bilateral lung bases. A clinical possibility of airway-esophageal fistula was considered and FB examination was performed.http://dx.doi.org/10.1155/2014/567582) On distal inspection, the wire mesh of the esophageal stent was seen over the posterior tracheal wall extending up to 1-2 cms above the carina. A diagnosis of tracheal erosion of the esophageal SEMS with TEF was confirmed. The patient was unfit for any surgical intervention and therefore bronchoscopic management was planned.FB demonstrated prolapse and erosion of the proximal end of esophageal stent through the posterior tracheal wall and formation of tracheoesophageal fistula (TEF) through which bubbling secretions was visualized.  was successfully deployed under flexible bronchoscopy and conscious sedation, to successfully cover the entire extent of the eroded esophageal SEMS and the TEF  and four months had passed since tracheal SEMS placement.Reported major complications associated with the placement of esophageal SEMS include significant bleeding, tumor obstruction, stent migration, airway-esophageal fistulization, gastroesophageal reflux, aspiration pneumonia, and persistent chest discomfort . EsophagDespite these factors, delayed tracheal erosion and TEF formation due to esophageal SEMS is a rarely described occurrence. However, erosion of esophageal SEMS into adjacent structures apart from the trachea such as aorta, bronchi, carotid arteries, and vertebral arteries has been described and vascular perforation is usually associated with a fatal outcome , 8. AggrThe most important principle of management in the situation of tracheal erosion by esophageal SEMS is to stabilize the airway and to seal off the esophageal-respiratory communication. The same may involve a combination of or either of the following steps: esophageal stent removal and/or repeat esophageal stent placement/coaxial esophageal SEMS/tracheal covered SEMS placement/surgery in cases of benign disease . BronchoA prophylactic approach for the prevention of this complications has also been suggested wherein parallel tracheal and esophageal stenting may be considered especially in situations where the large esophageal tumor burden increases the propensity of airway obstruction following esophageal SEMS deployment. In fact, in case a covered esophageal SEMS completely prolapses into trachea, fatal asphyxiation may occur which may not occur in case an uncovered stent had been utilized wherein the wire mesh can allow ventilation to continue .The present case highlights the fact that, in rare circumstances, esophageal SEMS can penetrate into the tracheobronchial tree and present with emergent and often life threatening airway obstruction or aspiration. Occasionally, short of a frank rupture into the trachea, the stent can cause compression effect and tracheal lumen compromise also and sometimes a limited penetration by a stent wire has also been reported . EndoscoVideo of the flexible bronchoscopy examination demonstrating the presence of eroded esophageal SEMS into the trachea. Covered metallic tracheal stent is being deployed using the flexible bronchoscope to cover the prolapsed esophageal SEMS.


Rice growth is severely affected by toxic concentrations of the nonessential heavy metal cadmium (Cd). To elucidate the molecular basis of the response to Cd stress, we performed mRNA sequencing of rice following our previous study on exposure to high concentrations of Cd . In this study, rice plants were hydroponically treated with low concentrations of Cd and approximately 211 million sequence reads were mapped onto the IRGSP-1.0 reference rice genome sequence. Many genes, including some identified under high Cd concentration exposure in our previous study, were found to be responsive to low Cd exposure, with an average of about 11,000 transcripts from each condition. However, genes expressed constitutively across the developmental course responded only slightly to low Cd concentrations, in contrast to their clear response to high Cd concentration, which causes fatal damage to rice seedlings according to phenotypic changes. The expression of metal ion transporter genes tended to correlate with Cd concentration, suggesting the potential of the RNA-Seq strategy to reveal novel Cd-responsive transporters by analyzing gene expression under different Cd concentrations. This study could help to develop novel strategies for improving tolerance to Cd exposure in rice and other cereal crops. Cadmium (Cd) is a widespread heavy metal pollutant that is highly toxic to living cells. Accumulation of the nonessential metal Cd in plants is a major agricultural problem. Specifically, Cd is absorbed by the roots from the soil and transported to the shoot, negatively affecting nutrient uptake and homeostasis in plants, even in very small amounts. Many agricultural soils have become contaminated with Cd through the use of phosphate fertilizers, sludge, and irrigation water containing Cd. Cd exposure inhibits root and shoot growth and ultimately reduces yield. Furthermore, Cd accumulation in the edible parts of plants such as seed grains places humans at a risk because of its highly toxic effects on human health. Reducing the Cd concentration in plants below the maximum level indicated by the Codex Alimentarius Commission of FAO/WHO  is necesCd causes oxidative stress and generates reactive oxygen species, which can cause damage in various ways such as reacting with DNA causing mutation, modifying protein side chains, and destroying phospholipids . VariousOryza sativa L. cv. Nipponbare) under a high Cd concentration using the RNA-Seq platform. A clear and detailed view of the transcriptomic changes triggered by Cd exposure is important to understand the gene expression network of the basal response to Cd stress. This could not be obtained from past studies using the microarray platform, but RNA-Seq can accurately quantify gene expression levels over a broad dynamic range with high resolution and sensitivity  ) . Enriched GO terms significantly in each cluster may represent the functional categories in rice under Cd exposure. Enriched GO terms of gradually upregulated transcripts under Cd exposure include metal ion transport (GO:0030001) , which may function in Cd transport. Response to oxidative stress (GO:0006979) and responsive to oxidative stress (GO:0006979) were also included in cluster 3 and cluster 4, respectively. This suggested that they might function in defense against Cd. Enriched GO terms of gradually downregulated transcripts under Cd exposure include translation (GO:0006412), translation elongation (GO:0006414), DNA replication (GO:0006260), and DNA repair (GO:0006281) . Photosynthesis, light harvesting (GO:0009765), and photosynthesis (GO:0015979) were also included in both clusters. These may function in plant growth. Thus, these correspond to the observed changes in phenotype  encoding iron-regulated transporter 1 (IRT1) was upregulated more than 5-fold under low Cd concentrations but responded only slightly under the high Cd concentration. IRT1s often transport Cd because of their low substrate specificity  that may function in Cd transport under Cd exposure. The expression of 183 transport transcripts was compared between low and high Cd concentration treatments in roots and shoots at 1 d, because Cd uptake from the hydroponic culture and efflux pumping are initial responses to Cd exposure (cificity –26, 30. We generated gene expression profiles for rice seedlings grown under low Cd concentrations. Phenotypic observations and constitutive gene expression indicated that low Cd concentrations cause growth retardation but are far from being fatal in rice. Several genes associated with defense systems were strongly upregulated; the expression of metal ion transporter genes tended to correlate with Cd concentration and GO enrichment analysis of the clustered genes based on their expression patterns, suggesting that our transcriptome profiles reflect responses to Cd in rice. Our data also suggest that it could be dangerous to eat plants that do not show specific Cd pollution symptoms growing in soil contaminated by small amounts of Cd. Establishing the exact composition and organization of the transcriptional network underlying the response to Cd exposure will provide a robust tool for improving crops in the future, for example, by creating low Cd uptake plants.Mapping of RNA-seq reads obtained from root and shoot samples to the reference IRGSP-1.0 genome sequence.


Immune-mediated pathogenesis has been suggested for idiopathic sensorineural hearing loss (iSNHL). Recent studies have investigated the relationship between iSNHL and autoantibodies against inner ear antigens, but specific tests are not available.To assess the positivity of autoantibodies against inner ear in a series of pediatric patients with idiopathic sensorineural hearing loss.We conducted a prospective, non-interventional observational study in a series of pediatric patients affected with iSNHL. Autoantibodies against inner ear , previously described in the serum of patients with Cogan's syndrome, were detected in our populations. The characteristics of children who resulted positive were also evaluated to verify if clinical data, disease progression and response to treatment could confirm an immune-mediated pathogenesis.Eleven patients were enrolled and 9 of them resulted positive for the inner ear antibodies detected. Non-organ specific autoantibodies were present in 5 children out of 9. An immune-mediated condition was diagnosed in 2 cases and minor immune manifestations were found in other 2 patients. In 5 cases hearing loss remained stable with no therapy, otherwise 4 children developed hearing loss progression. Two subjects were treated with steroids and methotrexate achieving hearing improvement. Another subject started methotrexate treatment showing hearing loss stabilization.Most of clinical characteristics and comorbidities described, added to immunologic tests results and to treatment efficacy, suggest an immune-mediated pathogenesis of hearing loss. Inner ear autoantibodies are not able to identify children affected with autoimmune sensorineural hearing loss but, integrated with clinical data, they can represent a diagnostic aid for the physician. Large prospective studies are needed to investigate usefulness, diagnostic and prognostic role of these autoantibodies.None declared.


Endovascular aneurysm repair (EVAR) is perhaps the most widely utilized surgical procedure for patients with large abdominal aortic aneurysms. This procedure is minimally invasive and reduces inpatient hospitalization requirements. The case involves a 72-year-old male who presented to the emergency department with right testicular ischemia two days following EVAR. Given the minimal inpatient hospitalization associated with this procedure, emergency physicians are likely to encounter associated complications. Ischemic and thromboembolic events following EVAR are extremely rare but require prompt vascular surgery intervention to minimize morbidity and mortality. Endovascular aneurysm repair (EVAR) is becoming the first-line treatment option for many patients with large abdominal aortic aneurysms (AAA) . IschemiA 72-year-old Caucasian male with a history of abdominal aortic aneurysm presented to the emergency department two days after abdominal aortic endovascular stent grafting. He presented with right flank, right lower abdominal, and right testicular pain. He reported persistent pain following surgery and was discharged from the hospital less than 24 hours prior to presentation on a bowel regimen and pain medication. Despite regular bowel movements and pain medication consumption, the pain persisted leading to his emergency department visit. He denied any recent fever, chills, headaches or vision changes, chest pain, shortness of breath, nausea, vomiting, dysuria, hematuria, or diarrhea. The pain is not improved with lying flat nor is the patient sexually active.On physical examination, his heart rate, blood pressure, respiratory rate, and temperature were within normal limits. Pertinent examination findings included a soft, exquisitely tender right testicle in vertical alignment with normal positioning and intact cremasteric reflex bilaterally. The pain persists despite testicular elevation. The abdomen was mildly distended with diffuse tenderness that was worse on the right. No peritoneal signs or costovertebral angle tenderness were noted. Femoral and dorsalis pedis pulses were present and equal bilaterally.Urology was consulted with coinciding history and physical exam findings. Following their clinical examination, urology recommended against performing emergent manual detorsion because the presentation was not consistent with testicular torsion. The patient subsequently underwent ultrasonographic evaluation  of the sIntermittent torsion was considered less likely. This was followed by a computed topographic angiogram (CTA) study , which rAn abdominal aortic aneurysm (AAA) is an abnormal dilation of the major abdominal artery found in 5% to 10% of men aged 65 to 79 years , 5. SeveSurgical repair of AAA can be attained via open surgical repair (OSR) or endovascular stent grafting known as endovascular aneurysm repair (EVAR) . EVAR isOf the known complications of EVAR, leakage of blood into the aneurysm sac (endoleak) is most common. Additional complications include stent migration, aneurysm sac expansion, stent occlusion, and delayed rupture , 6. IschIn a case report by McKenna et al., a man of similar age presented with acute onset left scrotal pain following EVAR . One notIn the case presented, acute ischemia was suspected clinically and confirmed ultrasonographically. Blood flow to the right testicle was reduced with right testicular decreased echogenicity following EVAR, without clinical signs of torsion or infarction. As opposed to the previously mentioned cases, this patient was managed conservatively with symptom control and close observation and required no surgical intervention. Repeat ultrasound demonstrated stable blood flow to the right testicle and symptoms had resolved prior to discharge.The exact pathophysiology of testicular ischemia and infarction following EVAR is not clear as this is a rare complication with scarce literature. McKenna et al.  report tEndovascular aneurysm repair is becoming the primary treatment option for many patients with operative AAA. Given the decreased intraoperative complications and reduced inpatient requirements, postoperative complications are more likely to occur at home and present to the emergency department for evaluation. These patients should be evaluated closely and in conjunction with vascular surgeons for potential end-organ damage secondary to acute ischemia and thromboembolic events.


Sustainable forest management is based on functional relationships between management actions, landscape conditions, and forest values. Changes in management practices make it fundamentally more difficult to study these relationships because the impacts of current practices are difficult to disentangle from the persistent influences of past practices. Within the Atlantic Northern Forest of Maine, U.S.A., forest policy and management practices changed abruptly in the early 1990s. During the 1970s-1980s, a severe insect outbreak stimulated salvage clearcutting of large contiguous tracts of spruce-fir forest. Following clearcut regulation in 1991, management practices shifted abruptly to near complete dependence on partial harvesting. Using a time series of Landsat satellite imagery (1973-2010) we assessed cumulative landscape change caused by these very different management regimes. We modeled predominant temporal patterns of harvesting and segmented a large study area into groups of landscape units with similar harvest histories. Time series of landscape composition and configuration metrics averaged within groups revealed differences in landscape dynamics caused by differences in management history. In some groups (24% of landscape units), salvage caused rapid loss and subdivision of intact mature forest. Persistent landscape change was created by large salvage clearcuts (often averaging > 100 ha) and conversion of spruce-fir to deciduous and mixed forest. In groups that were little affected by salvage (56% of landscape units), contemporary partial harvesting caused loss and subdivision of intact mature forest at even greater rates. Patch shape complexity and edge density reached high levels even where cumulative harvest area was relatively low. Contemporary practices introduced more numerous and much smaller patches of stand-replacing disturbance  and a correspondingly large amount of edge. Management regimes impacted different areas to different degrees, producing different trajectories of landscape change that should be recognized when studying the impact of policy and management practices on forest ecology. Forest policy and management practices within the U.S. have changed substantially following widespread dissatisfaction with management overly focused on the production of wood fiber and game species habitat. Over the past several decades, managers of public and private lands have to varying degrees incorporated a much wider set of objectives including the protection or provision of amenities, biodiversity, and ecosystem services ,2. Much The sustainable management of forest landscapes and development of effective forest policy requires an understanding of the functional relationships between management practices, changes in landscape conditions, and ecological response. Abrupt changes in forest policy or other drivers of landscape dynamics make it fundamentally more difficult to evaluate these relationships. Because ecological processes operate over a wide range of temporal scales, responses to landscape change are time-dependent. Changes in species presence or abundance are frequently delayed following periods of rapid landscape change, and ecological communities take time to equilibrate to new landscape dynamics imposed by new management practices –8. DelayEmpirical studies of forest loss or fragmentation effects commonly rely on a space-for-time substitution , where rSatellite images provide the synoptic views needed to characterize forest conditions and landscape change. The ~40-year depth of the Landsat image archive in particular facilitates studies of forest landscape dynamics. However, there are relatively few retrospective analyses of landscape dynamics following abrupt changes in forest management practices. In the Pacific Northwest region of the U.S., Landsat image time series have been used to address the consequences of federal forest policy change e.g., ,17). Lan. Lan17])Choristoneura fumiferana (Clem.)), a native pest that causes widespread defoliation and mortality of balsam fir  and spruce (Picea spp.) trees [The Atlantic Northern Forest of the northeastern U.S. encompasses roughly 11 million hectares within a transition zone between the northern boreal forest and the southern temperate deciduous-dominant forest. A substantial portion of this area lies within northern Maine, the largest contiguous block of undeveloped forestland in the nation (~4 Mha). Despite a long history of logging and commercial management for fiber production, major changes in management practices within recent decades have led to contemporary landscape conditions with little historical precedent. The spruce-fir forests of the region are subject to periodic infestations of the eastern spruce budworm (.) trees ,21. Main.) trees . Public .) trees  in 1989 .) trees  and less.) trees .Management practices in Maine have elicited concerns regarding the sustainable provision of forest values. During the budworm outbreak, salvage logging rates were well above recognized long-term allowable levels . RegenerThe objective of our research was to characterize predominant patterns of cumulative landscape change in the Atlantic Forest of northern Maine, and to evaluate how pre- and post-FPA management regimes have influenced landscape conditions across space and time. We used Landsat imagery and forest inventory data to develop and validate forest composition maps and a time series of forest harvest maps (1973–2010). We modeled predominant temporal patterns of harvesting and segmented a large study area into groups of landscape units with similar harvest histories. We then linked harvest history with changes in landscape composition and configuration in order to characterize the evolution of landscape conditions in response to forest management practices before and after abrupt change induced by the FPA. Our approach provided an objective synthesis of predominant patterns of change associated with specific landscape units, with the spatial and temporal resolution needed to attribute change to different management regimes.Acer rubrum, Acer saccharum, Betula alleghaniensis, Betula papyrifera, Fagus grandifolia) predominate across lower hilltops and at mid-slope. Spruce-fir species  predominate where soil or microclimatic conditions exclude the more demanding hardwoods. Mixedwood stands commonly occur along ecotones or as a result of successional dynamics following disturbance. Shade-intolerant hardwood species  are commonly found following intense disturbance. Periodic defoliation by spruce budworm is the most prominent form of natural disturbance. Windthrow is common but generally results in small canopy gaps [Our northern Maine, U.S.A. study region  was defiopy gaps . Virtualopy gaps  and rougForest harvest and composition maps were assembled from a time series of Landsat Multispectral Scanner (MSS), Thematic Mapper (TM), and Enhanced Thematic Mapper Plus (ETM+) images acquired during summer leaf-on conditions . ConsecuForest harvest maps were produced using a change detection procedure based on vegetation index values calculated from sequential Landsat images. As initially described by Sader and Winne , forest The improved sensitivity of the NDMI to partial canopy disturbance is generally attributable to the heightened sensitivity of mid-infrared wavelengths to differences in forest canopy structure, leaf area, and biomass ,35.We classified three-date NDMI and NDVI composites to produce forest change maps from TM/ETM+ and MSS image sequences, respectively. MSS imagery lacks a mid-infrared band required for calculation of the NDMI. This difference, coupled with reduced spatial and radiometric resolution, limits the efficacy of MSS imagery for detection of partial canopy disturbance. Disturbances mapped using MSS imagery (1973–1988) represent stand-replacing events, predominantly spruce budworm salvage clearcuts. Disturbances mapped using TM/ETM+ imagery (1988–2010) represent a wide range of intensities, and we differentiated two intensity classes interpreted as stand-replacing and partial canopy disturbance. The stand-replacing class was intended to represent harvests in which a new cohort was established following removal of a large proportion of the canopy, whether by clearcut as defined by the FPA ,37 or byHarvest maps were produced by unsupervised classification of overlapping three-date NDVI or NDMI image sequences . Classification of a three-date sequence mitigates the impact of cloud cover in the second image provided the first and third give clear views. An ISODATA algorithm applied to each three-date composite produced 50 statistical classes that were interpreted into forest disturbance, regrowth, and no-change information classes. Stand-replacing and partial harvest classes derived from TM/ETM+ imagery were differentiated based on the relative magnitude of NDMI change, guided by visual interpretation of Landsat imagery and available aerial photography. Confusion between light partial harvests and changes induced by factors such as atmospheric effects or interannual variability in forest phenology were resolved through on-screen editing . IndividWe produced a time series of maps depicting cumulative harvest impact (1975–2010) by overlaying successive harvest maps. For each time series date, a pixel was labeled as regenerating forest if preceding intervals included a harvest 1973–1988 or a stand-replacing harvest 1988–2010. A pixel was labeled as partially harvested if preceding intervals included only a single partial harvest. When preceding intervals included multiple partial harvests, pixels were labeled as regenerating forest, reflecting the anticipated ecological and silvicultural effects of multiple entries within the ~20-year period over which partial harvests were mapped (1988–2010). For each date of our time series, the result depicts the cumulative footprint of harvest operations since 1973.We mapped forest composition using equivalent unsupervised classification methods applied to each of the 1975 MSS and 2004 TM images. Dates were selected on the basis of cloud cover and image quality. For the purpose of forest type mapping, small areas of cloud cover in the 2004 image were replaced with data from the 2001 ETM+ image. Statistical classes produced from an ISODATA algorithm were aggregated to coniferous-dominant (>75% coniferous), deciduous-dominant (>75% deciduous), and mixed type classes through visual interpretation of Landsat imagery, with reference to available aerial photography and existing land cover maps. In some previously disturbed areas, exposed soils, woody debris, or herbaceous vegetation precluded the assignment of forest type and pixels were instead assigned to an indeterminate class. Patches less than 0.81 ha in size were removed and a 3x3 majority filter was applied to each map to consolidate patch boundaries and simplify the 2004 patch structure to more closely match the 1975 data.Assignment of ISODATA classes to forest types was subjective and sometimes difficult. A mistaken assignment could lead to bias in the representation of forest type extent. If for example pixels representing forest with a deciduous component of 70–75% were mistakenly committed to the deciduous-dominant class rather than the mixed class, the extent of the deciduous class would be overestimated according to the class definition of >75% deciduous. We used validation data obtained from field plots  Program provides quality-assured measurements of forest attributes from a national network of field plots adhering to a systematic sampling design . We madeFIA estimates of forest age have been used to validate Landsat-derived disturbance time series under the assumption that trees sampled for age estimation germinated at the time of disturbance . HoweverIn Maine, the contemporary FIA inventory design was established in 1999, with 20% of plots surveyed annually during sequential 5-year cycles. Although data are available from plots measured during earlier inventories, coordinate locations are known for only a fraction of those plots. We therefore used data collected during contemporary inventory cycles to discriminate past harvest intensity. A harvest event identified by image interpretation was labeled stand-replacing provided FIA age dated stand origin to 1970 or later  and field crews labeled the stand as either sapling or poletimber. However, for plots harvested after 1999, recorded stand age was an unreliable indicator of stand-replacing disturbance because age estimates frequently corresponded to a few residual stems rather than a newly established cohort. In these cases, intensity classes were discriminated using plot measurements made during consecutive 5-year inventory cycles; a harvest was labeled stand-replacing if plot basal area  was reduced by >70%.Our validation sample of 509 plots was insufficient to produce reasonably precise accuracy estimates for individual time series intervals. We therefore aggregated intervals into six validation classes: 1973–1988 stand-replacing harvest, 1988–1999 stand-replacing harvest, 1988–1999 partial harvest, 1999–2010 stand-replacing harvest, 1999–2010 partial harvest, and intact mature forest . Map and reference validation class labels were assigned in a manner consistent with the construction of cumulative harvest maps. Where multiple entries occurred, labels were assigned based on the date of first stand-replacing disturbance. Where multiple partial harvests occurred, labels of either 1988–1999 or 1999–2010 stand-replacing were assigned based on the date of second entry. Map and reference labels were compiled into an error matrix. Overall accuracy, user accuracy (the complement of class commission error), producer accuracy (the complement of class omission error), and corresponding standard error estimates were calculated by poststratification ,43. AddiThe 1975 and 2004 forest type maps were validated using FIA plot measurements of coniferous and deciduous live tree basal area collected during 1980–1982 and 1999–2003 inventories, respectively. Differences in dates between maps and field inventories were resolved by excluding samples where intervening harvests occurred. For 2004 map validation, we excluded plots mapped as harvested from 1999–2004; for 1975, we excluded plots mapped as harvested from 1975–1982. A sample of 445 plots remained for validation of the 2004 map; only 70 plots were available for validation of the 1975 map. As previously described, we identified coniferous-dominant and deciduous-dominant class thresholds for which errors were best balanced and mapped class extents least biased. Following refinements made to improve consistency between maps, an error matrix was compiled for each map based on selected threshold values. Estimates of overall, user, and producer accuracy were calculated by poststratification ,43.To quantify harvest rates through time, identify trends, and associate trends with changes in landscape conditions, we tessellated our study area into landscape units using a 5 km square grid . A 5 km TX and are equivalent to the loading vectors or principal components of a PCA. The spatial amplitudes correspond to the PCA scores obtained by projecting the time series of X onto the subspace spanned by the EOFs.An EOF analysis identifies a sequence of uncorrelated patterns or modes of variability that characterize variation within a two-dimensional data set . EOF anaA traditional EOF analysis or PCA is sensitive to extreme observations and outliers, which can distort the outcome such that dominant modes of variability represent contrasts between anomalous and regular observations rather than patterns of variability within regular observations . We perfPaired EOFs and amplitude functions comprise orthogonal modes of variability, ordered by the amount of total variance explained. We modeled cumulative harvest time series as linear combinations of dominant EOFs, selected based on the proportion of overall variance explained by successive modes. By including only dominant modes, modeled time series represent statistically coherent variability in harvesting patterns that occurred over large portions of the study area. The ROBPCA algorithm provides a measure of orthogonal distance between samples and the subspace defined by dominant EOF modes. Unusually large orthogonal distances indicate outlying samples that do not conform to characteristic patterns defined by dominant modes. We identified 12 orthogonal outliers using the ROBPCA nominal cutoff value  and exclTo classify predominant temporal patterns of harvesting from the EOF analysis and to associate those patterns with groups of grid cells, we performed an agglomerative hierarchical clustering  of modelLandscape composition metrics were calculated for grid cells and averaged within groups to evaluate changes associated with predominant harvesting trends. Within our time series of cumulative harvest maps, available forestland was classified as either regenerating, partially harvested, or intact mature forest . The EOF and cluster analyses produced time series of cumulative harvest area, the reciprocal of intact mature forest area. We also produced time series of cumulative partial harvest and regenerating forest area to evaluate changes in harvest intensity associated with predominant harvesting trends. Available forestland was summarized by 1975 forest type to associate harvest history and landscape change with initial landscape composition. To evaluate composition change as a legacy of harvest practices, we quantified forest type change between 1975 and 2004 for areas harvested before 2004. Composition change in unharvested forestland was not evaluated as part of this research.Early successional and intact late successional forest patches are landscape elements of particular interest, given the expansion of partial harvest practices in the 1990s. Cumulative changes in the patch configuration of regenerating and intact mature forest were evaluated by calculating time series of landscape metrics. We selected a small number of metrics of general relevance to forest ecology that quantify primary aspects of class configuration thought to have been affected by changes in management practices. Metrics were calculated using Fragstats version 4.2 . Area-weHarvest validation classes were mapped with an overall accuracy of 88% . User anForest type classes for 1975 and 2004 were mapped with overall accuracies of 76% and 68%, respectively Tables . User anBy 2010, 61% of available forestland was mapped as harvested, and 40% regenerated by stand-replacing or multiple partial harvests. Averaged across all grid cells, harvest rates increased ca. 1985 and then remained quite constant at about 2% per year  [Lepus americanus), are found at highest density within coniferous and mixed regenerating forest ~15–35 years post-harvest [Martes americana) [The changes in landscape composition and configuration we have quantified imply potentially important impacts on forest ecology and wildlife. Salvage clearcuts created large blocks of early successional forest habitat, benefitting the federally threatened Canada lynx (adensis) . In Main-harvest –59. The -harvest . Additioericana) . For speS1 AppendixDescription of image processing steps performed prior to forest harvest and composition mapping.(DOCX)Click here for additional data file.S2 AppendixDescription of the procedures used to validate cumulative forest harvest (1973–2010) and forest type maps (1975 and 2004), and a brief interpretation of validation outcomes.(DOCX)Click here for additional data file.


Fine needle aspiration (FNA) cytology is a well-established diagnostic method based on the microscopic interpretation of often scant cytological material; therefore, experience, good technique and smear quality are equally important in obtaining satisfactory results.We studied the use of fresh surgical pathology specimens for making so-called mock-FNA smears with the potential of cytohistological correlation. Additionally, we studied how this process aids the improvement of preparation technique and smear quality.Cytological aspirates from 32 fresh biopsy specimens from various sites: lung (20), lymph nodes (6), and breast (6) were obtained, all with a clinical diagnosis of tumor. Aspiration was performed from grossly palpable tumors. 25G needle and Cameco-type syringe holder was used with minimal or no suction.Unfixed surgical specimens provided sufficient cytological material that resulted in good quality smears. After standard processing of specimens into microscopic sections from paraffin embedded tissues, cytohistological case-series were created. No significant alteration was reported in tissue architecture on hematoxylin-eosin stained sections after the aspiration procedure. A gradual, but steady improvement was observed in smear quality just after a few preparations.Our study proved that surgical specimens may be used as a source of cytological material to create cytohistological correlation studies and also to improve FNA cytology skills. The use of very fine gauge needle  during the sampling process does not alter tissue architecture therefore the final histopathological diagnosis is not compromised. We conclude that by using fresh surgical specimens useful cytohistological collections can be created both as a teaching resource and as improving experience. Fine needle aspiration cytology or biopsy (FNAC/FNAB) is a well-established diagnostic method used in many countries as a first-line diagnostic procedure aiding preoperative management of various palpable and non-palpable lesions. The history of needle biopsy spans several centuries; however, its use as a routine diagnostic technique was established mostly during the twentieth century To accelerate the learning process and experience of recognizing specific cellular morphology we propose here the use of fresh (i.e. unfixed) surgical pathology specimens as a source of creating FNA smears and cytohistological correlation series. This procedure also might serve as a model for learning the basics of FNAC including sampling and smear preparation. Our main objective was to demonstrate how fresh surgical specimens may used as a source for cytological material with respect to creating cytohistological correlation case series. Additional objective was to study the usefulness of surgical pathology specimens in the practice of fine needle aspiration technique including sampling and smear preparation.The study was conducted in Mures County Emergency Clinic/Spitalul Clinic Judetean de Urgenta Mures, university hospital within the University of Medicine and Pharmacy of Tirgu Mures. Upon submission a signed written informed consent was obtained from all patients acknowledging that bioptic specimens removed by surgery due to therapeutic considerations might undergo additional studies. For conducting our study we did use only some parts of the bioptic specimens, without altering in any way the final histological diagnosis. Since no additional tissues/cells were removed from any patient, other than it would anyway be removed, approval of the ethics committee was not necessary. Samples presented in this study were not described nor published in other study, including parts or the whole material.The authors of this article did not interact with any of the patients whose surgical pathology specimens constituted the main subject of this article. The identity of patients was protected at several levels. All cases were designated with an ID number, starting with one. After the cyto-histological correlation series were created only these ID numbers identified each case. From clinical data only tumor localization, size, age, sex and histopathological type was linked to the identification numbers. This way patient confidentiality was respected by anonymizing bioptic specimens. The obtained cyto-histological correlation cases were archived according to case numbers  plus the above mentioned minimal clinical data, without any other data that could be linked to any individual .Fresh, unfixed, surgical biopsies, mostly whole-organs were used as the source for cytological material aspiration, prior formalin fixation. We obtained cytological aspirates from 32 fresh biopsy specimens from various sites: lung (20), lymph nodes (6), and breast (6). All organs were submitted to the pathology laboratory with a clinical diagnosis of tumor. Aspiration was performed exclusively from grossly palpable tumors. Cytological material was obtained using standard fine needle aspiration technique as described by Stanley A Cameco-type syringe holder mounted with a 10 ml three-piece syringe was used. For staining, we used two commercially available staining kits Cytocolor and Hemacolor  and standard Papanicolaou stain. In each case, we used the following two gauge type needles: 23G  and 25G .Aspired material was spread on standard pre-cleaned glass slides  using the standard one-step method described by Stanley and colleagues and its modification Most of the surgical specimens provided sufficient cellular quantity using the standard FNA technique. However, material obtained from lesions showing extensive desmoplasia or were rich in conjunctive stroma resulted in less cellularity. Nevertheless all cytological metrials obtained resulted in sufficient and mostly good quality smears. All the specimens were processed according to specific protocols and paraffin-embedded tissue blocks were created. Thus from all lesions microscopic slides were made and stained with hematoxylin-eosin. This permitted the creation of cytohistological correlation pairs. As depicted in There was no perceivable morphological differences between smears prepared with material aspired with 23G or 25G needles, both proved to be sufficent. In more fibrous or desmoplastic lesion 25G needles, handled too firmly, had a tendency to bend.The one-step method described by Stanley prooved to be optimal in preparing cytological material obtained from fresh speciemens Fine-needle aspiration cytology or biopsy (FNAC/FNAB) is increasingly performed as an economical, safe and rapid minimally invasive technique for diagnosis of various lesions including tumors and non-neoplastic lesions Using fresh surgical pathology specimens offers the possibility to experiment with various smearing and preparation techniques, the ultimate goal being to produce good quality smears. Before performing FNA at the patient’s side both dexterity and smear preparation skill may be improved by practicing on fresh surgical material without altering the histopathological interpretation of these specimens. Nevertheless there are few models described for practicing FNA technique, sampling and slide preparation. Some of these models include the use of fresh animal (beef and pork) liver introduced into a double layered latex glove, while others describe models made of silicone Finally, using surgical specimens also ensures that in addition to cytology, also paraffin-embedded tissue sections are also available at end of the day, enabling the development of the third foundation of cytodiagnosis, by the use of cytohistological correlation series see  and 2.We described our experience of using surgical pathology specimens (tissue biopsies) for practicing FNAB proficiency. Fresh surgical pathology specimens, especially whole-organs may be used for training residents prior to the beginning of FNAB performance on patients in the clinical setting. Surgical specimens may be used as a source of cytological material to improve FNAB skills. These specimens also provide cytological material to study preparation conditions that may alter smear quality. We found that fresh surgical pathology specimens used prior formalin-fixation were a good source of cytological material and may be used for practicing sampling and smear preparation.Additionally, using biopsy specimens offers the great opportunity to study cytohistological correlations. The creation of cytological smears has little impact on a laboratory’s financial expenses. No special equipment is needed and standard dyes (such as Papanicolaou and Romanowsky-types stains) may be used. For each specimen up to 3–4 cytological smears are optimal without significantly hindering grossing or delaying routine practice. Compared to standard histology additionally to glass-slides, stains and coverslips, only needles (with no larger gauge than 23–25G) and syringes  and fixative are needed that may be acquired at low cost. We consider that the costs of such materials are far beyond the benefits the introduction of mock-FNA into daily routine may represent.Aspired material can also be used to study and experiment with the various conditions that may critically alter smear quality and potentially hinder the formulation of the correct diagnosis. To become proficient in sampling and smearing, it is necessary to perform many biopsies on patients. However, the process can be significantly accelerated by performing practice sessions with fresh surgical pathology specimens before attempting the procedure in the clinical setting.


Healthy hearing depends on sensitive ears and adequate brain processing. Essential aspects of both hearing and cognition decline with advancing age, but it is largely unknown how one influences the other. The current standard measure of hearing, the pure-tone audiogram is not very cognitively demanding and does not predict well the most important yet challenging use of hearing, listening to speech in noisy environments. We analysed data from UK Biobank that asked 40–69 year olds about their hearing, and assessed their ability on tests of speech-in-noise hearing and cognition.About half a million volunteers were recruited through NHS registers. Respondents completed ‘whole-body’ testing in purpose-designed, community-based test centres across the UK. Objective hearing (spoken digit recognition in noise) and cognitive  data were analysed using logistic and multiple regression methods. Speech hearing in noise declined exponentially with age for both sexes from about 50 years, differing from previous audiogram data that showed a more linear decline from <40 years for men, and consistently less hearing loss for women. The decline in speech-in-noise hearing was especially dramatic among those with lower cognitive scores. Decreasing cognitive ability and increasing age were both independently associated with decreasing ability to hear speech-in-noise  among the population studied. Men subjectively reported up to 60% higher rates of difficulty hearing than women. Workplace noise history associated with difficulty in both subjective hearing and objective speech hearing in noise. Leisure noise history was associated with subjective, but not with objective difficulty hearing.Older people have declining cognitive processing ability associated with reduced ability to hear speech in noise, measured by recognition of recorded spoken digits. Subjective reports of hearing difficulty generally show a higher prevalence than objective measures, suggesting that current objective methods could be extended further. The DTT choice of single digit words promotes easy understanding and the opportunity to ‘fill-in’ missed acoustic information, while steady-state, speech-shaped noise enables masking across the speech range, but does not allow the ‘glimpsing’ that can reduce masking in more complex noise. Overall, DTT performance correlates more closely with PTA than do some other SiN tests Self-report of hearing difficulties, using questionnaire instruments or often only one or two questions, has commonly been used to assess the prevalence of hearing loss in large epidemiological studies In this study we investigated the relation between DTT SRT and cognitive ability as a function of age, sex, socioeconomic status, and noise exposure. The results were compared with self-report of hearing difficulties and findings from other studies, including some previously unpublished data from the NIH Toolbox www.ukbiobank.ac.uk.Complete details of all UK Biobank procedures are available at This research was covered by the UK Biobank ethics agreement. Within England, UK Biobank has approval from the North West Multi-centre Research Ethics Committee (MREC). All participants provided written informed consent.Participants , about one third of the participants  of cards showing 3 and 6 pairs of shapes, respectively. The shapes were concealed and the participant was required to recall locations of matching shape pairs. Correct responses were confirmed. The Reaction Time test of processing speed sequentially presented twelve pairs of shapes. The participant pressed a button as quickly as possible if a pair of shapes matched. Finally, in the Digit Span (‘Numeric memory’) test of verbal working memory participants were initially presented on the screen with two digits and were required to key in the digits in reverse order. After each correct response the digit sequence was increased by one. The task ended after two incorrect responses. Raw score was the maximum number of correctly recalled and ordered digits. Performance on the Prospective Memory, Reaction Time and Digit Span tests additionally depended on executive function and all tests depended on alerting and orienting attention.Variable numbers of participants completed each test , primariAnalysis of self-report measures used binary logistic regression. Standardised scores, which each maximised the variability within the sampled population, were derived for each cognitive test using principal components analysis. A composite score was defined as a simple sum of the five standardised scores. Both individual and composite cognitive scores were standardised to have zero mean and unit standard deviation, such that higher values corresponded to higher ability. Scores were grouped into deciles for analysis. Regression modelling techniques accounting for sex, socio-economic deprivation  tone detection thresholds declined from 18–85 years (NIH Toolbox data (DOCX)Click here for additional data file.


The physiological role of CNS regeneration inhibitors in the naïve, or uninjured, CNS remains less well understood, but has received growing attention in recent years and is the focus of this review. CNS regeneration inhibitors regulate myelin development and axon stability, consolidate neuronal structure shaped by experience, and limit activity-dependent modification of synaptic strength. Altered function of CNS regeneration inhibitors is associated with neuropsychiatric disorders, suggesting crucial roles in brain development and health.The growth inhibitory nature of injured adult mammalian central nervous system (CNS) tissue constitutes a major barrier to robust axonal outgrowth and functional recovery following trauma or disease. Prototypic CNS regeneration inhibitors are broadly expressed in the healthy and injured brain and spinal cord and include myelin-associated glycoprotein (MAG), the reticulon family member NogoA, oligodendrocyte myelin glycoprotein (OMgp), and chondroitin sulfate proteoglycans (CSPGs). These structurally diverse molecules strongly inhibit neurite outgrowth  Proper function of the adult mammalian CNS requires precise assembly, refinement, and maintenance of an elaborate network of neuronal connections. During development, axon guidance molecules facilitate the formation of intricate neuronal networks. After this initial assembly, many circuits are refined in an activity-dependent manner during a short time period of heightened plasticity, called the critical period (CP). At the end of the CP, the fine tuning of networks is complete and they acquire their mature shape. Synaptic contacts in the mature brain are stable over long time periods , display striking similarities to Mag null mice, suggesting that gangliosides may be the primary receptors responsible for MAG-mediated axon protection -anchor. OMgp is expressed by OLs and neurons in the CNS , are extracellular matrix (ECM) proteoglycans that consist of a protein core with covalently attached glycosaminoglycan (GAG) side chains , and its homolog RPTPσ  and long-term depression (LTD) of synaptic transmission are opposing forms of activity-dependent synaptic strength modifications, and are thought to underlie aspects of learning and memory formation   signaling. At the cell surface, BDNF binds TrkB to activate several growth promoting pathways, including mTOR complex 1 (mTORC1) and MAP kinase/ERK. However, CSPG binding to RPTPσ attenuates activity of TrkB (Kurihara and Yamashita, NgR1 null mice show impaired memory function, including impaired fear extinction and consolidation (Park et al., NgR1 and NogoA have been associated with schizophrenia (Sinibaldi et al., Many human neuropsychiatric disorders are associated with defects in synaptic structure or function and may be caused by a shift in excitatory/inhibitory balance (Pittenger, CNS regeneration inhibitors play important physiological roles in the uninjured brain and spinal cord. The myelin-associated inhibitors MAG, NogoA, and OMgp regulate myelin formation and axon-myelin interactions. NogoA, OMgp, and CSPGs regulate synapse formation and maturation, and they influence activity-dependent synaptic strength. Growing evidence suggests that some CNS regeneration inhibitors participate in an intricate cross-talk with growth promoting molecules at the level of several key signaling molecules. This finely tuned balance of excitation and inhibition in the developing and mature CNS may be necessary for proper formation and function of neural networks.The physiological role of these molecules raises important considerations for therapeutic strategies designed to promote neural regeneration following injury. The acquisition of a large number of ligand-receptor systems that restrict neural network plasticity may have been a prerequisite that enabled the evolution of larger and more powerful neural networks. Larger and more complex brains may be more vulnerable to aberrant changes in synaptic connectivity and therefore, once fully developed, need to be consolidated and protected by molecules that constrain excessive network rearrangement. Following injury to the adult CNS, molecules that restrict aberrant growth and plasticity may be detrimental since they limit attempts to modify or rebuild nearby networks to compensate for lost neural circuits. Therefore, a deeper understanding of the physiological roles played by CNS regeneration inhibitors is of great interest both clinically and biologically.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Local field potentials (LFPs), measured using extracellular electrodes, provide information about local neuronal population activity. The development of multi-electrode arrays (MEAs) that measure LFPs simultaneously at different network locations has led to a renewed interest in the LFP as a measure of neuronal network activity . While tin vitro.Here, we describe two contributions to the effort to understand LFP generation in neuronal networks. First, we investigated reduced compartmental models to assess their suitability for producing realistic spatial LFP characteristics. We show that the chosen reduction method  creates


ICR-CTSU introduced EDC in 2012 necessitating the development of EDC specific training materials for participating sites.Training of site staff on EDC systems is essential to ensure data quality and returns are maximised. Guidance notes are issued to support standardised completion of case report forms. However, due to the technical nature of EDC, guidance notes alone provide insufficient support for most site staff. Provision of effective, consistent and timely EDC training across a trial's lifespan is a continuing challenge, particularly for large trials or those with long follow-up where site staff turnover is an added consideration.ICR-CTSU created a custom EDC system user guide, developed training slides and included instructional text within the EDC system. Since the implementation of EDC, ICR-CTSU have developed short training videos covering key topics including logging in, record navigation, data entry and query response. The videos illustrate the EDC screen view and are accompanied by scripted voice over guidance. Training materials are accessible online via the ICR-CTSU website.Whilst customised EDC system user guides and in-built instructions form the basis of site staff training, supplementary technical training is required. This can be provided to site staff individually through webinars, teleconferences and centre initiation visits, however the online training videos provide a more consistent and accessible resource. This solution should reduce demand on ICR-CTSU resources for ongoing site training.


Ischaemic heart disease is most prevalent in the ageing population and often exists with other comorbidities; however the majority of laboratory research uses young, healthy animal models. Several recent workshops and focus meetings have highlighted the importance of using clinically relevant models to help aid translation to realistic patient populations. We have previously shown that mice over-expressing the creatine transporter (CrT-OE) have elevated intracellular creatine levels and are protected against ischaemia-reperfusion injury. Here we test whether elevating intracellular creatine levels retains a cardioprotective effect in the presence of common comorbidities and whether it is additive to protection afforded by hypothermic cardioplegia.+ cardioplegic arrest, as used during cardiac surgery and donor heart transplant, was further enhanced in prolonged ischaemia (90 minutes) in CrT-OE Langendorff perfused mouse hearts .CrT-OE mice and wild-type controls were subjected to transverse aortic constriction for two weeks to induce compensated left ventricular hypertrophy (LVH). Hearts were retrogradely perfused in Langendorff mode for 15 minutes, followed by 20 minutes ischaemia and 30 minutes reperfusion. CrT-OE hearts exhibited significantly improved functional recovery (Rate pressure product) during reperfusion compared to WT littermates . Aged CrT-OE mouse hearts (78±5 weeks) also had enhanced recovery following 15 minutes ischaemia . The cardioprotective effect of hypothermic high KThese observations in clinically relevant models further support the development of modulators of intracellular creatine content as a translatable strategy for cardiac protection against ischaemia-reperfusion injury. This was attributed to increased phosphocreatine (PCr) buffer capacity, elevated myocardial glycogen, improved ‘energy reserve’ and the ability of creatine to reduce the open probability of the mitochondrial permeability transition pore . CardiopRapid revascularization of ischaemic regions within the heart by thrombolysis or primary percutaneous coronary intervention remains at the clinical forefront to effectively treat AMI ,19. Pre-It should be noted that previous work from our laboratory has shown that very high myocardial [Cr] may itself cause LV hypertrophy and dysfunction , and defWe also performed experiments in mice aged 18 months since the ageing heart is known to have altered cardiac substrate metabolism , is more+ and Mg2+ ions to reduce metabolic demand and prolong ischaemic tolerance [ex vivo rat hearts [Cold, cardioplegic arrest of the heart remains the gold standard for effective myocardial protection during cardiac surgery  and hearolerance . In thisolerance  and in polerance  undergoit hearts . Howevert hearts . These bt hearts  and not ex vivo not in vivo experimental approaches and ideally we would have both. This reflects the limitations of having sufficient numbers of aged animals and the welfare implications of combining TAC surgery with ischaemia-reperfusion surgery. Furthermore, we did not perform tetrazolium staining in our ex vivo hearts since the period of reperfusion was insufficient for complete washout of NADH from necrotic tissue, which would lead to under-estimation of infarct sizes [During ischaemia, the heart undergoes many metabolic changes including a reduction in intracellular PCr . We did ct sizes . It is tct sizes .We have shown that elevating creatine levels in the mouse heart improves functional recovery following ischaemia-reperfusion even in the presence of old age or LVH. Both conditions previously associated with loss of cardioprotective efficacy. Furthermore, elevating intracellular creatine is additive to standard hypothermic cardioplegia for recovery from prolonged ischaemia. Together these findings support the development of small molecule activators that increase intracellular Cr , which wS1 File(DOC)Click here for additional data file.