We construct a state-and-transition model for mammals in tropical savannas in northern Australia to synthesize ecological knowledge and understand mammalian declines. We aimed to validate the existence of alternative mammal assemblage states similar to those in arid Australian grasslands, and to speculate on transition triggers. Based on the arid grassland model, we hypothesized that assemblages are partitioned across rainfall gradients and between substrates. We also predicted that assemblages typical of arid regions in boom periods would be prevalent in savannas with higher and more regular rainfall. Data from eight mammal surveys from the Kimberley region, Western Australia (1994 to 2011) were collated. Survey sites were partitioned across rainfall zones and habitats. Data allowed us to identify three assemblage states: State 0:- low numbers of mammals, State II:- dominated by omnivorous rodents and State III:- dominated by rodents and larger marsupials. Unlike arid grasslands, assemblage dominance by insectivorous dasyurids (State I) did not occur in savannas. Mammal assemblages were partitioned across rainfall zones and between substrates as predicted, but—unlike arid regions—were not related strongly to yearly rainfall. Mammal assemblage composition showed high regional stability, probably related to high annual rainfall and predictable wet season resource pulses. As a consequence, we speculate that perpetually booming assemblages in savannas allow top-down control of the ecosystem, with suppression of introduced cats by the dingo, the region's top predator. Under conditions of low or erratic productivity, imposed increasingly by intense fire regimes and introduced herbivore grazing, dingoes may not limit impacts of cats on native mammals. These interacting factors may explain contemporary declines of savanna mammals as well as historical declines in arid Australia. The cat-ecosystem productivity hypothesis raised here differs from the already-articulated cat-habitat structure hypothesis for mammal declines, and we suggest approaches for explicit testing of transition triggers for competing hypotheses. The processes that drive fluctuations in population size and community composition have long been a source of fascination for ecologists, and gaining a general understanding of their identity and effects remains an enduring goal. Although some emphasis has been placed on the influence of intrinsic factors, such as social interactions, in driving species' dynamics In many systems, the interplay between bottom-up and top-down forces can lead to the existence of alternative states that are characterized by different dominant species or species-groups State-and-transition models were initially developed to assist in managing non-equilibrial rangeland systems Here, we apply state-and-transition modelling to synthesize knowledge about native mammal assemblages in the Kimberley region of northern Australia and explore factors that might drive them. This region is of considerable conservation concern: populations of many mammal species are declining across the northern Australian savannas, conceivably towards irreversible regional or total extinctions. Worldwide, about a quarter of all mammal species are threatened with extinction; about 38% of land mammals suffer from habitat loss associated with diversion of natural resources to benefit humans, and 16% are at risk from hunting or harvesting Although mammal declines elsewhere in Australia have received considerable attention Sminthopsis macroura, Pseudomys desertor, Rattus villosissimus, Mus musculus) and functional groups (particularly insectivorous dasyurids and omnivorous rodents).We use a recently described model from central Australia There are two key differences between central Australian arid grassland assemblages and those of the Kimberley's tropical savannas: (1) The former are now largely devoid of medium-sized mammals (e.g. peramelids) whereas all medium-sized mammals persist in the tropical savannas, albeit in most cases, with greatly reduced ranges It is important that state-and-transition models be testable annual rainfall,rocky versus non-rocky savannas, andthe abundance and representation of particular species and taxonomic or functional groups.Because of the importance of antecedent rainfall in driving patterns of mammalian abundance and composition through time, we predicted also that inter-annual differences in rainfall would lead to shifts between states. Rainfall, and thus productivity, is predictable in the Kimberley but not in central Australia, where erratic rainfall drives ephemeral pulses of productivity. In consequence, we predicted further that Kimberley mammal assemblages would be more stable and exist in higher states than those in central Australia. We use our results to generate hypotheses about the key factors that prompt shifts between assemblage states and hence further our understanding of the threats to the Kimberley mammal faunas. These hypotheses also can be extended to, and tested in, other tropical savanna areas.4 grasses dominating the ground layer The Kimberley region is in the far north west of Australia and represents the Western Australian component of Torresian  habitats. It has a tropical monsoonal climate with rain falling mostly during the summer from November to April. Annual rainfall ranges from 1400 mm on the far north coast to <400 mm along the semi-arid southern boundary . AlthougTo classify mammal assemblage states, we collated data from 229 survey sites in savanna across all the major Kimberley bioregions All survey data presented here consist of one-off site surveys. Where repeat site surveys occurred, we maintained temporal independence by choosing just one survey at random to represent that site. As different rainfall regions and savanna habitats provide the potential conditions needed to drive different states and transitions, we considered this space-for-time approach to be appropriate.This study makes use of data from eight separate mammal fauna surveys with differing methodologies and trap effort per site . DespiteAnalyses were performed on individual species and also on pre-defined functional groups , terrestrial omnivorous rodents (Muridae), arboreal rodents, large marsupials (>150 g) including dasyurid predators, bandicoots (peramelids), possums , and small macropods (macropodids <2000 g)). We compared species and taxonomic/functional groups in terms of their abundance, measured here as percentage trap success  for each survey. We tested for potential under-estimation of small dasyurids among surveys by separately analyzing trap records from pitfall traps based on the rationale that pitfall traps are the most effective method for estimating the abundance of small dasyurids in arid grassland surveys The hypothesis that species and mammal groups differ in abundance among rainfall zones and between rocky and non-rocky savannas was tested using Kruskal-Wallis tests. This non-parametric test was chosen due to inequality of sample sizes and non-normality of the survey data, which had many zero values. The effect of fluctuations in annual rainfall within rainfall zones/savanna types  was tested using regressions of trap success over previous wet season rainfall. Differences among mammal assemblages across rainfall zones and savanna habitats were explored using Bray Curtis ordinations, with the Sorensen dissimilarity distance used to calculate separation between surveys. A non-parametric multi-response permutation procedure (MRPP) was used to test for differences among predefined rainfall zone and savanna habitat (rocky and non-rocky) groups. We used PC-ORD 4 for ordinations and MINITAB 14 for all statistical analyses.Zyzomys argurus, Pseudomys nanus, Rattus tunneyi and P. delicatulus. The next most prominent group comprised large dasyurid predators , with 0.95% trap success. Small insectivorous dasyurids were relatively rare and yielded a total trap success of only 0.09%. Other groups represented in the surveys included bandicoots (0.58%) and arboreal rodents (0.43%). The low trap success rate for macropods and possums (<0.2%) probably reflects ineffective sampling rather than low numbers as these animals are not often trapped even where locally numerous.After removal of data from sites sampled more than once, we recorded a total of 5138 mammals from 71,016 trap nights (mean trap success 7.24%) . The larCanis lupus dingo and feral cats Felis catus were abundant. Dingoes were frequently observed and recorded in the high and medium rainfall zones during surveys . Similarly, cats are present throughout the Kimberley Although not recorded by trapping, dingoes P. delicatulus, P. nanus) were more numerous in low rainfall zones and non-rocky savannas, while others  were most abundant in high rainfall rocky savannas  showed variable responses to rainfall and habitat. Some rodent species , the bandicoot Isoodon auratus and arboreal rodents occurred only in surveys in the high rainfall zone , including rodents and small dasyurids, were captured using pitfall traps irrespective of rainfall zone . RodentsIn individual surveys, 17% of sites had no mammals (State 0), 2% were dominated by insectivorous dasyurids (State I), 37% were dominated by omnivorous rodents (State II) and 44% had a significant component of large mammals in addition to omnivorous rodents (State III). Surveys yielding zero mammals (State 0) were unusual in high (8% of surveys) and medium (0%) rainfall rocky savannas but more common in non-rocky savannas  and in low rainfall rocky savannas (32%). Small dasyurids (State I) dominated assemblages in 1% of high rainfall rocky savannas, and in 5% and 10% of rocky and non-rocky sites in the low rainfall zone. Rodents dominated assemblages (State II) in medium rainfall non-rocky (73%), low rainfall rocky (63%) and low rainfall non-rocky (71%) savannas. State II assemblages were also common in medium rainfall rocky (25%) and high rainfall non-rocky savannas (38%), but less so (6%) in high rainfall rocky savannas. State III assemblages with large CWR mammals (>150 g) were restricted to high rainfall or rocky savannas, with 84% and 75% of surveys yielding these assemblages in high and medium rainfall rocky savannas, and 40% in high rainfall non-rocky savannas.Z. argurus and P. desertor and on non-rocky substrates (sand plains and open non-rocky savannas) by P. nanus, P. delicatulus and R. tunneyi. These species differ from their arid zone counterparts where State II assemblages are dominated by Notomys alexis and Pseudomys hermannsburgensis (Letnic and Dickman 2010). Finally, surveys in the high rainfall zone conformed to State III arid zone assemblages, with rodents and marsupial groups including predatory dasyurids dominating. State III assemblages were dominated by rodents  and the large dasyurid predator Dasyurus hallucatus on rocky substrates. State III assemblages also included bandicoots (Isoodon macrourus and I. auratus), arboreal rodents (Mesembriomys macrurus and Conilurus penicillatus) and infrequently trapped possums (e.g. Wyulda squamicaudata) and small macropods . The latter groups, comprising CWR mammals, have become extinct since European settlement in central Australian grasslands This study confirms the existence of alternative assemblage states for mammals in tropical savannas of the Kimberley region. A proposed model depicting these states is shown in Sminthopsis macroura, S. virginiae, Planigale ingrami, P. maculata and Pseudantechinus ningbing occurred throughout Kimberley. However, they never occurred at high abundance, nor were they ever more common than other mammal groups. In contrast, arid assemblages are often dominated by small dasyurids (i.e. State I) A key difference between the arid zone assemblages described by Letnic and Dickman D. hallucatus, P. tapoafata), bandicoots and arboreal rodents have generally been restricted to high rainfall savannas in the Kimberley and Top End regions since the 1980s The assemblages described here correspond closely with previous reports on mammals in the Kimberley savannas Key transition triggers for changes in mammal assemblage state in central Australian grasslands are rainfall events that drive ephemeral pulses of primary production, and wildfires, drought, grazing by introduced herbivores and predation by introduced predators (cats and foxes) D. hallucatus) Despite the predictability of these patterns, assemblage states were not fixed; for example, State II assemblages sometimes occurred in the high rainfall zone and State III assemblages in the medium rainfall zone. Several other studies have also documented changes in assemblage states at particular sites over time High regional stability among assemblages of Kimberley savanna mammals in response to inter-annual differences in rainfall suggests that transitions there are governed differently from those in central Australian grasslands. Because savanna productivity is generally relatively high due to higher rainfall than in more arid regions, factors that effect downward state transitions may assume more importance in driving assemblage changes. Such factors operating in the savannas may also explain the relatively low abundance of mammals in most Kimberley assemblages relative to those in the arid grasslands during periods of population irruption Predation by introduced cats and foxes drives mammal assemblages towards lower states in arid regions Of the predators in the Kimberley savannas, dingoes are most likely to have a top-predator role and to influence mammal assemblage states. Ecological theory and many empirical observations show that where resource pulses are regular, as in the Kimberley relative to the arid zone, predators often maintain higher and more stable populations and can then exert strong regulatory effects on prey If dingoes act as ‘biodiversity regulators’ As in the conceptual model for arid Australian assemblages, fire regimes and introduced herbivores have major influences on assemblages of savanna mammals Arid zone mammals typically occur as State 0 or State I assemblages for long periods until shifted up by large but infrequent flooding rains The cat-ecosystem productivity hypothesis posited here, and depicted in


It has been previously shown that loss of consciousness is associated with a breakdown of dominating fronto-parietal feedback connectivity as assessed by electroencephalogram (EEG) recordings. Structure and strength of network connectivity may change over time. Aim of the current study is to investigate cortico-cortical connectivity at different time intervals during consciousness and unconsciousness. For this purpose, EEG symbolic transfer entropy (STEn) was calculated to indicate cortico-cortical information transfer at different transfer times.The study was performed in 15 male volunteers. 29-channel EEG was recorded during consciousness and propofol-induced unconsciousness. EEG data were analyzed by STEn, which quantifies intensity and directionality of the mutual information flow between two EEG channels. STEn was computed over fronto-parietal channel pair combinations  to analyze changes of intercortical directional connectivity. Feedback  and feedforward  connectivity was calculated for transfer times from 25 ms to 250 ms in 5 ms steps. Transfer times leading to maximum directed interaction were identified to detect changes of cortical information transfer  induced by unconsciousness (p<0.05).The current analyses show that fronto-parietal connectivity is a non-static phenomenon. Maximum detected interaction occurs at decreased transfer times during propofol-induced unconsciousness . Strength of maximum feedback interaction decreases during unconsciousness (p = 0.026), while no effect of propofol was observed on feedforward interaction. During both consciousness and unconsciousness, intensity of fronto-parietal interaction fluctuates with increasing transfer times.Non-stationarity of directional connectivity may play a functional role for cortical network communication as it shows characteristic changes during propofol-induced unconsciousness. Altered states of consciousness are associated with changes in directional (effective) and functional connectivity within a fronto-parietal network. This is observed as a common phenomenon during anesthesia, vegetative states and sleep Recent studies show that functional connectivity of cerebral areas is not a static phenomenon, but shows spontaneous fluctuations over time However, so far, fluctuations of neuronal network connectivity have been shown for functional connectivity as measured by functional magnetic resonance imaging (fMRI). The current study evaluates whether directional cortical connectivity as measured by electroencephalogram (EEG) may also show fluctuations, but in the range of milliseconds Fifteen healthy male volunteers  participated after approval by the university's ethics committee  and informed written consent. The study was conducted in the department of anesthesiology of the Technische Universität München, Germany. Standard monitoring parameters and 29-channel EEG  were continuously recorded with a sampling rate of 5 kHz using BrainAmp  electroencephalographic amplifier and BrainVision Recorder  data acquisition software. Propofol was administered by a target controlled infusion pump . Subjects received propofol in stepwise increasing concentrations until loss of consciousness occurred, which was defined clinically when subjects stopped following the investigator's instruction to squeeze his hand. TCI concentration at this point was maintained for 10 minutes to allow equilibration. EEG was recorded without drug application  and during propofol-induced unconsciousness (10 minutes). Basic artifact detection  and average reference were performed on EEG recordings.In existing EEG data of propofol-induced unconsciousness we applied EEG analysis within the framework of an information-theoretic approach allowing a model-free exploration of directed interactions between frontal (process X) and parietal (process Y) electrodes. For this purpose, EEG symbolic transfer entropy (STEn) was calculated For calculation, STEn analyzes consecutive sequences For frontal (x) and parietal (y) EEG this indicates feedforward parietal → frontal projections between parietal and frontal electrodes . STEn was computed over fronto-parietal channel pair combinations at consciousness and propofol-induced unconsciousness using one artifact free EEG signal of 10 s length for each subject . Acording to the signal length criterion Values of STEn may indicate spurious interdependencies in the EEG caused by spectral changes. To address this question, surrogates from the electroencephalographic signals were derived by maintaining the spectral amplitudes  and randomising the phase  non-parametric STEn showed consistent results even in different setups, e.g. embedding parameters, electrode montage Further studies with different anesthetics and altered states of consciousness are required to clarify some open points. Unconsciousness was induced by propofol. Therefore, it remains an open question, whether the detected results are a propofol-specific phenomenon or reflect characteristics of unconsciousness in general. The small number of subjects and low spatial resolution of the fronto-parietal network analyzed in EEG represent further limitations. Method inherent issues can not be fully excluded. Estimation of transfer times as performed in the present analysis may also include intrinsic prediction within one area In conclusion, our results indicate that cortical information transfer is not a static process but fluctuates periodically. Fluctuations of directional connectivity seem to reflect an intrinsic phenomenon of the brain to energetically optimize cortical communication. Characteristically, fluctuations are changed during propofol-induced unconsciousness towards faster information transfer. At the same time, the amount of parieto-frontal feedforward projections is not reduced during unconsciousness, while fronto-parietal feedback interactions are reduced. Both the inhibition of feedback connectivity and the altered fluctuation of information transfer during propofol administration are proposed as functional correlates of loss of consciousness.


Quality of Life (QoL) data from oncology trials may have missing data which cannot be assumed to be missing completely at random (MCAR) . IgnorinWe searched MEDLINE (2002-2012) to identify oncology trials reporting longitudinal analysis of QOL data. The appropriateness of the analysis was reviewed and trials reporting QOL as primary/secondary endpoint were assessed for reporting quality using the CONSORT extension for PROs .69 RCTs reporting longitudinal QOL analyses were identified. 29 (42%) use an analysis to account for the nature of the missing data. Methods varied widely, eg pattern-mixture models, conditional linear models, QTWiST, joint longitudinal models, generalised estimating equations, selection models and Markov models. Fourteen papers used more than one method check the robustness of their results.In order for QOL data to adequately inform clinical decision-making the correct analysis needs to be performed. Statistical methods ignoring the missing data were found to over-estimate QOL but it was rare for the significance of QOL differences between treatments to change. A strategy for appropriate analysis of QOL data will be presented using case studies to highlight where ignoring informative missing data could alter the conclusions regarding treatment differences.


Progressive increase of temperatures as well as longer seasonal drought periods revealed by climate studies correspond to fast environmental changes that forest species face with their actual genetic background. Natural selective processes cannot develop an adaptive response within this time frame. Thus the capability of forest tree species to adapt to the new environments will depend on their genetic background, but also rely on their phenotypic plasticity. Several reports have shown the involvement of epigenetic modifiers as the basis of the phenotypic plasticity, and in particular to the adaptation to abiotic stresses. DNA methylation (methylation of cytosine residues)is one the most important epigenetic modification in eukaryotes. Itis involved in specific biological processes such as gene transcription regulation, gene silencing, mobile element control or genome imprinting.Therefore, there is a great interest in analyzing cytosine methylation levels and distribution within the genome.HpaII and MspI are isoschizomers that are frequently used to detect cytosine methylation. HpaII cannot cleave if one or both cytosines are fully methylated (in both strands), whereas MspI cleaves C5mCGG but not 5mCCGG sequences). For each sample, MSAP analysis is performed using both EcoRI/HpaII and EcoRI/MspI digested samples. Comparative analysis between EcoRI/HpaII and EcoRI/MspI fragment patterns allows the identification of two types of polymorphisms: (1) “Methylation-insensitive polymorphisms” that are associated with genetic variability and will show common EcoRI/HpaII and EcoRI/MspI profiles among samples; and (2) “Methylation-sensitive polymorphisms” that are associated with epigenetic variability and detected as amplified fragments differing in their presence or absence or in their intensity between EcoRI/HpaII and EcoRI/MspI patterns of the same sample. Thus, full methylation of the internal cytosine at the assayed CCGG sites will be associated with fragments detected in EcoRI/MspI pattern which are absent or less intense in EcoRI/HpaII profile. Hemi-methylation of the external cytosine will be associated with fragments observed in EcoRI/HpaII pattern which are absent in EcoRI/MspI profile.In order to analyze methylation-sensitive anonymous CCGG restriction sites we used MSAP technique (Methylation-Sensitive Amplified Polymorphism), an AFLP-based technique for the analysis of cytosine methylation. The technique is based on the use of isoschizomers that show differential cleavage sensitivity to cytosine methylation. Fagus sp.) and gymnosperm (Pinus sp.) species.We have optimized DNA methylation analysis for two forest tree species, including both angiosperm (Pinus pinea L. a specie which is characterized by a low genetic variability and high level of phenotypic plasticity. Representative populations spanning the whole distribution of this specie within Spanish geography were chosen for this study, including coastal and interior populations. MSAP patterns from vegetative propagated trees were compared among and within populations. Percentage of methylation-sensitive polymorphisms was calculated and selective markers were identified.The set-up of the MSAP technology has allowed study intra-specific variability of DNA methylation in Fagus sylvatica L. in response to water deficit we analyzed two populations of beech from Spain and Sweden which differ in the response to water stress. Patterns of DNA methylation clearly differed between populations. Although the rate of DNA methylation was similar between irrigated and non-irrigated trees in each polpulation after AMOVA, some specific MSAPs associated with water stress response were identified.To test a possible role of DNA methylation in the plasticity of


DNase I hypersensitivity (DHS) combined with next generation sequencing (DNase-seq) is an efficient way of observing, in a single experiment, the genome-wide chromatin effects associated with the binding of multiple transcription factors. Using quantitative contrasts of DHS before and after estrogen and androgen stimulation in breast and prostate cancer cell lines, we have shown that differential DHS can accurately predict hormone induced transcription factor binding. Despite its effectiveness, the DHS assay can vary significantly depending on the experimental parameters. To increase the robustness of this assay, we have systematically evaluated two major parameters, digestion level and fragment size. We found that while there is a broad range of suitable digestion level, over-digestion dramatically decreases the efficiency of detecting DHS regions. More interestingly, we found that different fragment sizes capture distinct chromatin elements, and thus represent different chromatin structures. We were able to classify different combinations of estrogen receptor coregulators that resulted in different local chromatin structures.


Aging is worldwide recognized as a dominant risk factor for most forms of cardiovascular disease -3. HowevIn adult and elderly humans, high levels of physical activity preserve the protective role of preinfarction angina against in-hospital mortality and cardiogenic shock after myocardial infarction. More recent data indicate that higher levels of physical activity performed before primary coronary angioplasty may independently predict the reduction of early and late cardiac mortality in older infarcted patients . Accordi


Mitotic chromosome motions have recently been correlated with electrostatic forces, but a lingering "molecular cell biology" paradigm persists, proposing binding and release proteins or molecular geometries for force generation.Pole-facing kinetochore plates manifest positive charges and interact with negatively charged microtubule ends providing the motive force for poleward chromosome motions by classical electrostatics. This conceptual scheme explains dynamic tracking/coupling of kinetochores to microtubules and the simultaneous depolymerization of kinetochore microtubules as poleward force is generated.We question here why cells would prefer complex molecular mechanisms to move chromosomes when direct electrostatic interactions between known bound charge distributions can accomplish the same task much more simply. Molecular mechanisms underlying mitosis, particularly those associated with directed chromosome movement during the cell cycle, have been pursued intensely over the past two decades with no clear picture emerging--or is there? Recent experiments identify positively charged kinetochore-associated molecules  that likely interact with negatively charged microtubule ends to generate electrostatic-dependent poleward forces that drive chromosome motion . This coIndeed, current thought on mitotic motions is shifting from a molecular to a more electrostatics-based framework -3, and psic] attempted to explain chromosome motions by nanoscale electrostatics and unnecessary sophistry..." [Perhaps the most surprising part of this story is the untimely resistance to classical electrostatics by the cell biology community. For example, critiques including "...groundless speculations in which the authors [stry..." , and reqstry..."  suggest stry..." ,2, and istry..."  and redustry..." .To gain perspective on this subject, it may be instructive to consider the problem of cell division in an evolutionary context, and more specifically in an ancestral cell that lacked "modern" molecular machinery. Clearly, cells have been dividing since the origin of life, and the mechanisms underlying this fundamental process in modern cells are likely derived from some ancestral state--just like other cellular processes  were likely derived from ancestral, catalytic RNAs that were later supplemented with supporting proteins. In a simple cell, all chromosome movements during mitosis are readily explained by electrostatic interactions between core components of the system , without the requirement for supplemental protein machinery -6. Why tOur current bottleneck in understanding mitotic chromosome movements seems reminiscent of another challenging question in our imperfect scientific history, namely the self-imposed constraints of ancient Greek astronomers in trying to explain geocentric planetary motions with perfect circles. Indeed, layers of epicycles were incorporated into an increasingly complex scheme of integrated circles that was "understood" by only the best natural philosophers of the time. It took ~2,000 years of scientific work by Brahe, Galileo, Kepler and Newton to achieve the simplicity of a modern theory based on a different conceptual scheme, i.e., elliptical orbits in a heliocentric solar system.Twenty years ago, Guenter Albrecht-Buehler lamented the view of many cell biologists that "molecular analysis of cellular functions" is the only acceptable approach to cell biology , yet thiThe authors declare that they have no competing interests.DHS made intellectual contributions and drafted the manuscript. LJG conceived the study. All authors read and approved the final manuscript.


We conducted a qualitative study to gain insight into the high retention rate and variable outcomes achieved in the 3-year PODOSA (Prevention of Diabetes and Obesity in South Asians) family-based cluster randomised controlled trial of a dietitian-delivered lifestyle modification intervention.We interviewed 21 trial participants and family volunteers following trial completion. Purposive sampling ensured representation of the diversity within the trial population. Data were thematically analysed. Findings were shared with trial dietiticans once the trial had closed.Many participants were aware of their family history of diabetes and attracted by the availability of information and monitoring. Offering home or clinic-based interventions, communication in the participant's chosen language(s) and excellent relationships between participants and dieticians contributed to retention. Adaptations in food choices were accommodated by participants, although community and faith based considerations made consistent adherence challenging. Participants found increasing their level of physical activity difficult given other demands, including long working hours, physically demanding employment and domestic commitments.Undertaking qualitative research allowed in-depth insight into participation, retention and adherence in the PODOSA trial. The main benefit participants sought was information regarding diabetes. Home-based interventions and continuity in dieticians were included in trial design, but it was not anticipated that these relationships would contribute so greatly to retention. Adaptations in food choices were not felt to be an extra burden upon participants but were made more difficult due to cultural considerations. Increasing levels of physical activity were felt to be an additional burden, again augmented by cultural considerations.


Rodent models of heterotopic cardiac transplantation are important for gaining a better understanding of heart allograft rejection. This study compared a working heart (WkHt) transplant model, which have developed by our laboratory, with the traditional non-working heart (Non-WkHt) model using cardiac MRI (CMRI) and pathological evaluation. Our results indicate that the physiological hemodynamic loading significantly impacts graft immune cell infiltration and myocardial function. Our WkHt transplant model with intact pulmonary circulation, retains hemodynamic loading close to that of native heart, and is thus more appropriate for both functional and immunological studies, especially for developing the clinically relevant CMRI techniques for comprehensive evaluation of the heart graft.The traditional heterotopic heart transplantation rodent model ,2 lacks WkHt model: En bloc of graft heart and lung was transplanted into the recipient abdomen. Graft aorta and superior vena cava (SVC) were anastomosed with recipient abdominal aorta and inferior vena cava (IVC), respectively.,4 Non-WkAlthough isograft hearts from both models had strong heart beats and normal ECG, they exhibited very different cardiac functions revealed by CMRI Fig.. The WkHOur results indicate that hemodymamic loading significantly impacts the graft status. The WkHt transplant model, with intact pulmonary circulation, retains physiological conditions close to that of the native heart, and therefore is more appropriate for functional and immunological studies, especially for developing the clinically relevant CMRI techniques for comprehensive evaluation of the heart graft.This study is supported by grants from the National Institutes of Health (P41EB001977 and R01HL-081349).


Oxaliplatin in combination with 5-fluorouracil (5-FU) and leucovorin (FOLFOX) is a common chemotherapeutic regimen for advanced colorectal cancer. Here, we present a case of interstitial lung disease associated with FOLFOX therapy.A 74-year-old man with a history of metastatic colorectal cancer was admitted with a four week history of progressive dyspnoea and evidence of severe respiratory failure. He had recently completed six cycles of FOLFOX chemotherapy in the months prior to presentation. Investigations did not reveal convincing evidence of infection or pulmonary embolism. CT chest demonstrated widespread pulmonary infiltrates and interlobular septal thickening. The patient was commenced on both broad spectrum antibiotic therapy and high dose corticosteroid treatment however his respiratory failure continued to progress. The patient died four days after admission due to progressive respiratory failure. Subsequent post-mortem examination demonstrated evidence of diffuse alveolar damage without evidence of tumour infiltration, infection or pulmonary embolism.Although infrequent, pulmonary toxicity can occur in association with FOLFOX therapy. Cessation of therapy and prompt initiation of corticosteroids may improve outcomes. Oxaliplatin in combination with 5-fluorouracil (5-FU) and leucovorin (FOLFOX) is an effective chemotherapeutic regimen for advanced colorectal cancer, improving survival with acceptable tolerability and side effect profile.A 74-year-old man with a history of metastatic colorectal cancer was admitted with a four week history of progressive dyspnoea and haemoptysis. He had undergone a right hemicolectomy three and a half years previously for a Duke’s A colorectal cancer. Three years post resection, a large single liver metastasis was identified with no other evidence of metastatic disease on PET scanning. CT examination of the chest at the time demonstrated only mild emphysematous changes .The patient was commenced on FOLFOX chemotherapy  and underwent six cycles from October to December 2008. A re-staging CT scan in January 2009 demonstrated progressive malignant disease with multiple new metastases within the liver. Chest imaging at this time revealed bilateral peripheral subpleural reticular changes and interlobular septal thickening without evidence of overt lung metastases . No chanThe patient subsequently presented in early February 2009 with four weeks of worsening dyspnoea and haemoptysis. His Creactive protein level was moderately elevated although white cell count and differential were normal. Arterial blood gases demonstrated severe type I respiratory failure. CT pulmonary angiogram excluded pulmonary embolism but demonstrated new patchy infiltrates with interlobular septal thickening . Sputum He was commenced on empirical broad spectrum antibiotics and high dose intravenous corticosteroid therapy. His respiratory failure worsened and after discussion with both the patient and his family, mechanical ventilation was not administered. The patient died four days after admission from progressive respiratory failure.Post-mortem examination did not demonstrate evidence of an infective aetiology on staining and culture of fresh specimens. There was also no evidence of emboli, thrombus or malignant infiltration within the lungs. Histological examination demonstrated widespread generalised changes within both lungs, with oedema, intra-alveolar haemorrhage and fibrin deposition along with interstitial and intraluminal fibroblastic proliferation without mature fibrosis. These findings were consistent with diffuse alveolar damage.Diffuse alveolar damage (DAD) is a common histopathological finding in patients with ARDS and is commonly associated with infections (both respiratory and systemic). It is more frequently seen in immunocompromised patients. Although drug reactions are a relatively uncommon cause of DAD, chemotherapeutic agents are overrepresented as potential precipitants. Mortality following histologically proven DAD obtained from surgical lung biopsy has been reported to be greater than 50%.7et al. demonstrated improved response rate, median time to progression and overall survival in patients with advanced colorectal cancer treated with FOLFOX in comparison to a regimen utilising Irinotecan instead of Oxaliplatin.17Oxaliplatin, in combination with 5-fluorouracil and leucovorin (FOLFOX) has been shown to be an effective regimen for the treatment of advanced colorectal cancer.Although pulmonary toxicity from FOLFOX appears to be an uncommon and apparently idiosyncratic effect of therapy, this case highlights the importance of clinicians being aware of this potentially fatal complication and the importance of regular follow-up.Given the temporal relationship between the use of FOLFOX and the onset of radiological changes and clinical symptoms, we believe that FOLFOX is the likely precipitant in this case. Whilst this appears to be an uncommon adverse effect, early recognition may allow prompt cessation of FOLFOX therapy and initiation of systemic corticosteroids, and this has the potential to improve outcomes for such patients.


Batrachochytrium dendrobatidis - Bd) are major drivers of amphibian declines worldwide. Habitat loss regulates host-pathogen interactions by altering biotic and abiotic factors directly linked to both host and pathogen fitness. Therefore, studies investigating the links between natural vegetation and chytridiomycosis require integrative approaches to control for the multitude of possible interactions of biological and environmental variables in spatial epidemiology. In this study, we quantified Bd infection dynamics across a gradient of natural vegetation and microclimates, looking for causal associations between vegetation cover, multiple microclimatic variables, and pathogen prevalence and infection intensity. To minimize the effects of host diversity in our analyses, we sampled amphibian populations in the Adirondack Mountains of New York State, a region with relatively high single-host dominance. We sampled permanent ponds for anurans, focusing on populations of the habitat generalist frog Lithobates clamitans, and recorded various biotic and abiotic factors that potentially affect host-pathogen interactions: natural vegetation, canopy density, water temperature, and host population and community attributes. We screened for important explanatory variables of Bd infections and used path analyses to statistically test for the strength of cascading effects linking vegetation cover, microclimate, and Bd parameters. We found that canopy density, natural vegetation, and daily average water temperature were the best predictors of Bd. High canopy density resulted in lower water temperature, which in turn predicted higher Bd prevalence and infection intensity. Our results confirm that microclimatic shifts arising from changes in natural vegetation play an important role in Bd spatial epidemiology, with areas of closed canopy favoring Bd. Given increasing rates of anthropogenic habitat modification and the resulting declines in temperate and tropical frogs, understanding how vegetation cover and disease interact is critical for predicting Bd spread and developing appropriate management tools for wild populations.Habitat loss and chytridiomycosis (a disease caused by the chytrid fungus  Anthropogenically driven habitat change has important implications for host-pathogen interactions, because even slight changes in environmental conditions can modify numerous biotic and abiotic factors that influence these interactions Batrachochytrium dendrobatidis (Bd), for instance, is more prevalent and occurs at higher infection intensities in pristine tropical forests compared to disturbed habitats Bd-induced amphibian declines Shifts in microclimate and changes in host community structure across gradients of habitat alteration play important roles in amphibian epidemiology Bd, thus limiting pathogen growth and persistence Bd persistence in these environments From the host's perspective, immune responses usually decrease as a result of the multiple effects of habitat alteration Bd  in populations of the common Green Frog (Lithobates clamitans) across a gradient of natural vegetation and microclimate. We sampled frogs in the southern Adirondack Park, New York State, a region with relatively low amphibian diversity and high dominance of this habitat generalist host Bd in amphibians and (ii) test for causal associations linking vegetation cover and microclimate with both Bd prevalence and infection intensity. Combined, these goals may elucidate whether vegetation and microclimate modulate disease risk in temperate amphibian populations affected by anthropogenic habitat change.Here, we examined the infection dynamics of Bd is enzootic and widespread in the northeastern U.S. Bd growth in this region Lithobates clamitans, Ranidae), the locally dominant amphibian host species. Green Frogs breed in permanent ponds during the boreal spring and summer, and typically spend most of the time at the shallow banks of water bodies −1. We recorded body weight (g) for each captured individual and screened post-metamorphic frogs with sterile swabs to quantify Bd prevalence and infection intensity (average of 16.6 L. clamitans per site). We tested samples for Bd in singlicate using Taqman qPCR Bd in each sample. This protocol maximizes amplification efficiencies by diluting extracts to reduce inhibition in environmental samples. For calculations of Bd prevalence, we categorized individuals as Bd-positive when their qPCR showed an infection load of greater than or equal to one g.e. Bd prevalence as the percentage of infected individuals and Bd infection intensity as average number of g.e. per population.We sampled anuran populations in the Adirondack Park of the State of New York . This region is heavily forested with elements representative of temperate and boreal forests −1) as a proxy for host population density. We specifically chose this study system because of its relatively low species diversity and high single host species dominance. These community attributes allow us to test hypothesis about the roles of microclimate in disease dynamics without the potential effects of complex host community structure. Nonetheless, we recorded host community diversity , and we reported the model with the highest goodness-of-fit for each run. We also used CAR to test for associations of natural vegetation with host population  and community attributes .To control for the effects of spatial autocorrelation among ponds, we analyzed our data using conditional autoregressions (CAR). We used CAR to test the relationship of each explanatory variable with Bd infection parameters. Because canopy density may be a better proxy for microclimate than our actual temperature records from a single data logger per pond, we tested an alternative path diagram in which canopy density directly affected Bd infection parameters. We compared goodness-of-fit among models using Expected Cross-validation Index (ECVI), an AIC-based index. We conducted CAR using Spatial Analysis in Macroecology v4.0 We used path analyses to statistically test for the strength of unidirectional cascading effects linking natural vegetation, canopy density, water temperature, and Bd at all study sites with mean prevalence of 24.25%±16.41 SD and mean infection intensities of 29.36±151.60 SD, reaching a maximum load of 1063.23 g.e. in our focal species without observed mortalities. Canopy density was the best predictor of Bd prevalence in L. clamitans , maximum daily average temp. , minimum daily average temp. ], and natural vegetation . We found no significant relationships between Bd prevalence and average body weight, capture rate, elevation, host community diversity, or overall host community biomass.We detected Bd infection intensity. Canopy density best predicted Bd infection intensities , maximum daily average temp. , minimum daily average temp. ], natural vegetation , average body weight , and capture rate  were also significant predictors of Bd infection intensity in L. clamitans. Similar to prevalence, we found no direct associations of Bd infection intensity with elevation, host community diversity, or overall host community biomass.We found similar results for Bd prevalence and infection intensity, our model selection identified three key environmental factors: canopy density, natural vegetation, and daily average water temperature  and infection intensity  underscores the dominance of the focal species across this landscape . The amount of natural vegetation surrounding our sampling ponds did not significantly predict host population and community attributes; namely host average body weight , host capture rate , overall host community biomass , species richness , and host diversity .The high evenness and low diversity among our sampling ponds . Although these species may help maintain Bd throughout the cold months, it is unlikely that they have a strong effect on disease dynamics later in the season, because they spend long periods of time away from water bodies Bd growth Bd infections in the wild Bd prevalence and infection intensity in temperate amphibian populations by modulating shade and associated microclimatic patterns, and that host community structure plays at most a minor role in our study system.The effects of habitat loss and forest fragmentation on host-pathogen interactions are varied Bd prevalence and infection intensity were environmental variables associated with vegetation cover and microclimate  in Pennsylvania showed that the proportion of leaf-litter and vegetation in the pond substrate correlated positively with Bd prevalence and infection intensity Bd transmission by providing substrates for Bd growth. Alternatively, leaf-litter may be an indicator of canopy cover and total degree of shade, which are potentially the true drivers of higher Bd levels in natural forest habitats. Similarly, in Costa Rica and Australia, localities with little to no canopy cover may provide amphibians with a refuge from Bd-induced extinction Bd year round (above 69%) Bd infections by environmental factors is a general phenomenon, and that these environmental controls linking natural vegetation to Bd dynamics are similar in tropical and temperate amphibian communities.The best predictors of mate see . The amontensity . A studyBd growth and persistence in both host and the environment, and (ii) by changing host's ability to fight and clear Bd through thermoregulation. The growth rate of Bd in the laboratory is strongly temperature-dependent, with an optimum climatic envelope ranging between 17–25°C and reduced persistence at temperatures above 28°C Bd growth may have been limited during some periods in our highest-temperature ponds. In fact, in four of our ponds 40.85±14.42% SD of water temperature records were above 25°C during the study period. Our data closely match the association between Bd infections and temperature found by Raffel et al. Bd infection intensities and prevalence at sites with average water temperatures above 23°C. In the State of Maine, USA, Bd infections were lower during the peak of the summer than in spring and fall, presumably because water temperatures often exceeded the optimal temperature range for Bd growth Bd-infected frogs were largely absent from sites where spring water exceeded 25°C Bd strain showed that pathogen growth in vitro is hampered by short exposures  to high temperatures that would typically be available to frogs at basking sites Bd infections Bd infection and transmission in open habitats Bd loads Bd infection intensity and frog survival were unrelated to water temperature Microclimate can affect amphibian disease risk in two ways by (i) regulating Bd system are larger than the effects imposed by density-dependent forces that typically predict prevalence and infection intensity in other temperate amphibians Bd infection intensity, potentially indicating density-dependent controls; however, this effect was marginal and capture rate was not a significant predictor of Bd when considered together with environmental factors in model selection. The potential effect of density on Bd was not linked to forest cover because we did not detect a significant effect of natural vegetation on host capture rate. Pathogen build-up to lethal infection intensities is more likely to occur in dense populations, under conditions that promote continuous reinfection of the hosts Our results indicate that the small-scale effects of vegetation and microclimate on our host-Bd susceptibility Bd prevalence and infection intensities. One potential explanation is that disease risk drops with age in response to host-acquired immunity, as repeated exposure to a given pathogen increases host resistance Bd infection intensity when considered independently, this parameter became a weak predictor when considering other environmental variables in the analysis. This weak effect of host body weight on Bd infection coupled with the fact that vegetation cover had no influence on host capture rate is an indication that, in our study system, habitat change has a larger influence on pathogen fitness than on host fitness. This result suggests that our focal species is highly resistant to Bd regardless of microclimate and vegetation. In fact, L. clamitans persists with a local Bd strain in the laboratory within optimal Bd grow temperatures Earlier studies in both tropical and temperate zones have found ontogenetic differences in Bd infections in both temperate and tropical systems Bd epidemics Bd resistance or tolerance could be a useful tool for assisted reintroductions in the wild Bd is most prevalent. With the high rate of anthropogenic modification to temperate and tropical forests, understanding how vegetation cover and disease interact is critical for predicting Bd spread and developing appropriate management tools for wild populations. Our results indicate that species-specific in situ management strategies will need to consider fine-scale microclimatic factors to safeguard Bd-susceptible species with narrow geographic distributions We have shown that disturbances to natural forest habitats reduce Table S1Model selection for environmental and biological variables influencing Batrachochytrium dendrobatidis (Bd) prevalence and infection intensity in populations of Lithobates clamitans in the Adirondack region, New York, USA.(DOCX)Click here for additional data file.


This study intends to develop a physiologic thumbprint for nonconvulsive seizures (NCSz) after acute brain injury. Abnormal electrographic brain activity including NCSz is common after acute brain injury and is associated with poor outcome. Mechanisms underlying this phenomenon are poorly understood but in animals periods of inadequate perfusion during seizures have been documented. In the present study we hope to gain better understanding of the relationship between abnormal electrographic patterns and brain homeostasis in patients with subarachnoid hemorrhage (SAH).2), regional cerebral blood flow (rCBF), and intracranial pressure monitoring; 69% (n = 36) also with intracortical EEG . Each minute of EEG  was categorized separately into non-ictal, on the ictal-interictal continuum (including periodic discharges at 2 Hz or faster), or seizures. We identified seizure onsets on ICE recordings and extracted the physiologic monitoring data 30 minutes pre and post seizure onset. Physiologic profiles based on standard error of the means plots were generated using high-frequency time series physiologic measurements and interpreted by visual analysis.Between June 2006 and June 2010, 51 poor-grade SAH patients underwent multimodality monitoring with microdialysis, brain oxygen tension  of patients with ICE recordings . NCSz were preceded by an increase in rCBF starting 15 minutes prior to onset of depth NCSz that stayed elevated throughout the observation period. Heart rate, mean arterial, intracranial, and cerebral perfusion pressures were elevated surrounding NCSZ. There was a small transient drop in PbtOThese findings confirm in comatose human beings that NCSz detected by ICE are associated with hyperemia, increased metabolism, and possibly brain tissue hypoxia, which serve as surrogates for secondary brain injury. Future research should implement novel approaches for ICU time-series data analysis, evaluate surface seizures, and utilize other surrogates of brain metabolism such as microdialysis.


Groin infections resulting in arterial bleeding due to bacterial vessel destruction are a severe challenge in vascular surgery. Patients with them most often present as emergencies and therefore need individualized reconstruction solutions.Case 1 is a 67-year-old man with infectious bleeding after an autologous reconstruction of the femoral bifurcation with greater saphenous vein due to infection of a bovine pericard patch after thrombendarterectomy. Case 2 is a 35-year-old male drug addict and had severe femoral bleeding and infection after repeated intravenous and intra-arterial substance abuse. Both patients were treated with an autologous obturator bypass of the superficial femoral vein. We review the current literature and highlight our therapeutic concept of this clinical entity.Treatment should include systemic antibiotic medication, surgical control of the infectious site, revascularization and soft tissue repair. An extra-anatomical obturator bypass with autologous superficial femoral vein should be considered as the safest revascularization procedure in infections caused by highly pathogenic bacteria. Severe groin infections with inguinal blood vessel destruction may be caused by intravenous substance abuse, radiation scars, transfemoral interventions and, most commonly, infection of prosthetic vascular implants (incidence 2% to 18%). ComplicStaphylococcus aureus (MRSA) superinfection of an inguinal lymphatic fistula. For surgical control of bleeding, orthotopic revascularization with iliacofemoral and iliacoprofundal greater saphenous vein interposition and resection of the xenogenic patch material was performed, accompanied by systemic MRSA-specific antibiotic treatment. Three weeks later rebleeding occurred due to an infectious arterial pseudoaneurysm. Intra-operative findings revealed complete erosion of the profundal venous graft anastomosis  or severe claudication-7. Witho% or seveThe obturator bypass, with primary patency rates of up to 76% after 2 years, has proven to be feasible in a few small clinical series with a considerable risk of late prosthetic graft infection remaining,3,4,9-11Streptococcus in this report. With proven susceptibility of several native and prosthetic materials to MRSA and rising clinical incidence, femoral vein obturator bypass is the safest revascularization procedure with the best long-term results[To the best of our best knowledge, investigators of only one clinical series have reported the use of obturator bypass fashioned from autologous superficial femoral vein at infection sites. We ther results.Pseudomonas aeruginosa and Group B Streptococcus. It should be considered the primary treatment option by vascular surgeons confronted with this problem, especially in young patients.The extra-anatomic obturator bypass with femoral vein is a safe and feasible revascularization procedure in patients with severe groin infections and highly pathogenic bacteria such as MRSA, Written informed consent was obtained from the patients for publication of this case report and any accompanying images. A copy of the written consent is available for review by the Editor-in-Chief of this journal.The authors declare that they have no competing interests and did not receive any payment from industrial partners.RK, CB, CT and AB performed surgery on the patients and were responsible for postoperative intensive medicine care. UL performed vascular graft infection studies in mice and was a major contributor in writing the manuscript. All authors read and approved the final manuscript.


This case study followed one adolescent patient who underwent bilateral deep brain stimulation of the centromedian parafascicular complex (CM-Pf) for debilitating, treatment refractory Tourette's syndrome for a period of 1.5 years. Neurocognitive testing showed no significant changes between baseline and follow-up assessments. Psychiatric assessment revealed positive outcomes in overall adaptive functioning and reduction in psychotropic medication load in this patient. Furthermore, despite significant baseline psychiatric comorbidity, this patient reported no suicidal ideation following electrode implantation. Deep brain stimulation is increasingly being used in children and adolescents. This case reports on the positive neurologic and neuropsychiatric outcome of an adolescent male with bilateral CM-Pf stimulation. Tourette syndrome (TS) is a chronic, childhood onset neuropsychiatric disorder, which occurs in approximately one percent of the general population, consists of motor tics and at least one vocal tic lasting longer than one year . Tic symHigh-frequency (above 100 Hz) deep brain stimulation (DBS) is being increasingly used as an efficacious treatment modality in patients with severe TS complicated by behavioral and psychiatric comorbidity that is refractory to first and second line treatment . Here, wA 17-year-old left-handed male with severe Tourette's syndrome, attention deficit hyperactivity disorder (ADHD), and obsessive compulsive disorder (OCD) was evaluated by pediatric neuropsychology and child psychiatry prior to the date of electrode implantation for DBS for treatment of refractory tic symptoms. The patient was treated with bilateral centromedian parafascicular complex (CM-Pf) stimulation and was followed for 1.5 years. Preoperative neuropsychometric testing was noteworthy for borderline general intellectual functioning and commensurate academic achievement . CognitiPreoperative psychiatric evaluation demonstrated severe psychosocial impairment related to treatment refractory Tourette's syndrome. Social impairment was associated with significant amounts of missed school and anxiety related to tics. Attempts to garner employment were hampered with being overwhelmed by his neurologic symptoms. Past psychiatric history was significant for three prior psychiatric hospitalizations, all occurring within the context of managing a complicated psychotropic medication regimen, and treatment approach for comorbid psychiatric conditions . InpatiePostoperative neuropsychometric testing revealed general cognitive functioning that had remained stable across time . The patPostoperative psychiatric evaluation was noteworthy for significant functional and psychosocial gains. No concerns for suicidal or self-injurious thoughts or actions were noted. The patient's psychotropic medication load was also significantly reduced . ImproveThis case report focused on the neurocognitive and psychosocial outcomes in an adolescent male who had received DBS for severe refractory Tourette's syndrome followed for 1.5 years. This patient demonstrated no significant changes in his follow-up psychometric testing and reported significant improvement in psychosocial functioning postoperatively. He was able to decrease his psychotropic medication load during follow-up psychiatric assessments and did not report any suicidal ideation throughout the course of treatment. DBS is being increasingly used for treatment-refractory TS in adults and is an important emerging treatment modality of treatment-refractory TS within the pediatric population . Thus faCase reports documenting long-term treatment outcomes in both adults and adolescents with severe refractory TS using DBS have yielded mixed results with regards to improved psychosocial functioning and overall quality of life. Stimulation of the globus pallidus internus (GPi) was shown to be beneficial in one adolescent boy with severe TS and comorbid OCD and anxiety but had little therapeutic benefit in another patient with similar tic severity, but lacking comorbid psychiatric conditions , 7. AckeIn a larger prospective cohort study, Porta and colleagues documented improvement in tic symptoms, as well as improvement in neuropsychiatric symptoms without cognitive decline in a 24-month followup of 15/18 patients who underwent bilateral thalamic stimulation [Given the ongoing need for awareness of cognitive and psychiatric outcomes in adolescent patients who have undergone DBS treatment, we specifically focused on the neurocognitive and psychosocial outcomes in an adolescent male who had received DBS for severe refractory TS . This ca


Acute variceal bleeding continues to be associated with significant mortality. Current standard of care combines hemodynamic stabilization, antibiotic prophylaxis, pharmacological agents, and endoscopic treatment. Rescue therapies using balloon tamponade or transjugular intrahepatic portosystemic shunt are implemented when first-line therapy fails. Rescue therapies have many limitations and are contraindicated in some cases. Placement of fully covered self-expandable metallic stent is a promising therapeutic technique that can be used to control bleeding in cases of refractory esophageal bleeding as an alternative to balloon tamponade. These stents can be left in place for as long as two weeks, allowing for improvement in liver function and institution of a more definitive treatment. Acute variceal bleeding continues to be associated with significant mortality. Recently published randomized controlled trials have shown that mortality from acute variceal bleeding has decreased over the past two decades from 42% to 15%, but this figure is still remarkably high [Balloon tamponade (BT) using Sengstaken-Blakemore tube used to be the primary therapy prior to the availability of endoscopic methods. Sengstaken-Blakemore tube, was first described in 1950 , is a muSurgical management for bleeding esophageal varices falls into two categories: shunt and nonshunt procedures. Nonshunt procedures include esophageal transection and gastroesophageal devascularization. Shunt procedure is classified into selective shunts such as splenorenal shunts, partial shunts such as calibrated small-diameter portacaval shunts, and nonselective shunts such as portacaval shunts. Surgical procedures are used less frequently in the era of advanced endoscopic therapy, TIPS, and liver transplant. However, surgical intervention remains an important and effective treatment modality in selected patients . Distal TIPS is an intrahepatic shunt that is placed using a percutaneous approach. It connects the hepatic vein and intrahepatic branch of portal vein using an expandable metallic stent with a diameter of 8 to 12 mm. TIPS was introduced as an alternative to surgery in the 1990s and has replaced surgical shunts in most centers . CurrentThe previously listed limitations of the current rescue therapies have led to the continued search for other methods as rescue therapy for refractory esophageal variceal bleeding.Self-expandable metal stents (SEMSs) are increasingly used in treatment of esophageal obstruction, stricture, leak, perforation, and tracheoesophageal fistula . AnecdotThe initial pilot study by Hubmann et al. in 2006 reported the use of SEMS in 20 patients with massive ongoing esophageal variceal bleeding . All patHubmann et al. published their extended series of 39 patients with massive ongoing esophageal variceal bleeding despite prior use of endoscopic or pharmacological therapy . ResultsWright et al. reported their experience using the SX-Ella Danis stent in ten patients with variceal bleeding and contraindications to TIPS insertion or balloon tamponade . The patDechêne et al. recently reported their experience using SEMS  in 8 patients with refractory esophageal variceal bleeding events . One patStandard fully covered SEMSs have also been used successfully for the management of esophageal tears caused by Sengstaken-Blakemore tube or banding. These applications further confirm the important role of SEMS in the successful management of iatrogenic esophageal injuries , 26, 27.Limited data suggests that specially designed SEMS (SX-Ella Danis stent) can effectively stop refractory bleeding from esophageal varices . This stStent placement requires appropriate training and expertise. Gastric varices will not be adequately compressed by the stent and persistent variceal bleeding after stent placement should raise the suspicion for presence of bleeding gastric varices. Appropriate precautions to prevent aspiration are needed since the stent is positioned at the gastroesophageal junction. Distal stent migration into the stomach was observed frequently but was not associated with apparent complications. Stent-related compression of the left main bronchus was reported in one patient and was treated successfully with stent removal , 28. No Current rescue therapies for bleeding esophageal varices are effective in stopping the bleeding in the majority of patients . In some


Pragmatic trials of treatment pathways can require patient awareness of treatment allocation in order to better represent clinical reality, for example when concordance with a treatment has an important effect on outcome. Conversely, masking of treating clinicians to allocation group can be impossible when full clinical assessment requires knowledge of the current treatments, resources do not permit separate teams for treatment and assessment or when such duplication of clinician contact might affect an outcome such as patient experience.The LiGHT trial is a 718 subject multi-centre 6-year NIHR-funded study of two treatment pathways for glaucoma with outcome measures of health related quality of life and cost effectiveness. We aimed to minimise variation in aspects of clinical behaviour that might introduce bias by affecting either of these outcomes.Custom-written decision support software permits real-time decision-making using analysis of multiple clinical measures made by masked observers: optic disc analysis, visual field assessment and intra-ocular pressure measurements. Treatment targets are objectively defined according to disease severity criteria. Target attainment accounts for known measurement uncertainties.A treatment algorithm based on robust, published criteria determines clinical decisions that influence cost or patient experience .Where objective criteria for clinical decisions exist, it is possible to balance the competing demands of reducing bias with protocol-driven escalation criteria with a pragmatic ‘real-world’ approach to therapeutic choices permitting variation in clinical practice, in a manner acceptable to clinicians recruiting into an unmasked pragmatic trial.


Picea rubens) populations at high elevation sites in Southern Appalachians to anthropogenic sulfate depositions. Red spruce seedlings are more sensitive to drought and cold stresses elicited by exposure to anthropogenic sulphate air pollution, than old trees. Genetic variation in seedlings and young trees was significantly reduced in heavily polluted stands. Several candidate genes involved in cold acclimation and calcium metabolism demonstrated signatures of selection corresponding with sulfate pollution levels. SNP allele frequencies at one gene involved in calcium metabolism demonstrated directional selection in response to anthropogenic sulfate deposition in red spruce growing at severely polluted high elevation sites, which corresponds well with the putative role of this gene in adaptation to acidification stress. Unlike range-wide experimental designs (e.g. the popular FST outlier test) and nucleotide diversity-based association studies, our within-population testing approach disentangled the confounding effects of geographic variation and demography from the genetic effects of recent selection.Rapid environmental changes, such as anthropogenic air pollution, can create significant evolutionary pressures on populations, species and ecosystems. Evolutionary processes occurring in natural populations at very short time scales, especially in response to human-induced environmental changes, are not well understood. Confounding effects of geographic variation and demography cannot be easily separated from signatures of recent selection in natural populations. We investigated the genetic response of declining red spruce (


The goal of enhanced nutrition in critically ill patients is to improve outcome by reducing lean tissue wasting. However, such effect has not been proven. This study aimed to assess the effect of early administration of parenteral nutrition (PN) on muscle volume and composition by repeated quantitative computer tomography (qCT).We performed a preplanned substudy of a randomized controlled trial (EPaNIC) that compared early initiation of PN when enteral nutrition was insufficient (early PN) with tolerating a pronounced nutritional deficit for 1 week in the ICU (late PN) [Critical illness evoked substantial loss of femoral muscle volume in 1 week, irrespective of the nutritional regimen. Early PN reduced the quality of the muscle tissue, as reflected by the attenuation, revealing increased intramuscular water/lipid content. Early PN also increased the volume of adipose tissue islets within the femoral muscle compartment. These changes in skeletal muscle integrity correlated with caloric intake. In the abdominal muscle compartments, changes were similar, albeit smaller. Femoral and abdominal subcutaneous adipose tissue compartments were unaffected by disease and nutritional strategy.Early PN did not prevent the pronounced wasting of skeletal muscle observed over the first week of critical illness. Moreover, early PN increased the amount of adipose tissue within the muscle compartments.


Varanus komodoensis). The growth of 400 individual Komodo dragons was measured in a capture-mark-recapture study at ten sites on four islands in eastern Indonesia, from 2002 to 2010. Generalized Additive Mixed Models (GAMMs) and information-theoretic methods were used to examine how growth rates varied with size, age and sex, and across and within islands in relation to site-specific prey availability, lizard population density and inbreeding coefficients. Growth trajectories differed significantly with size and between sexes, indicating different energy allocation tactics and overall costs associated with reproduction. This leads to disparities in maximum body sizes and longevity. Spatial variation in growth was strongly supported by a curvilinear density-dependent growth model with highest growth rates occurring at intermediate population densities. Sex-specific trade-offs in growth underpin key differences in Komodo dragon life-history including evidence for high costs of reproduction in females. Further, inverse density-dependent growth may have profound effects on individual and population level processes that influence the demography of this species.Somatic growth patterns represent a major component of organismal fitness and may vary among sexes and populations due to genetic and environmental processes leading to profound differences in life-history and demography. This study considered the ontogenic, sex-specific and spatial dynamics of somatic growth patterns in ten populations of the world’s largest lizard the Komodo dragon ( Somatic growth patterns represent a major component of an organism’s life-history Sex-specific growth variation must occur to result in sexual size dimorphism (SSD), reflecting different tactics and requirements for energy acquisition and investment between maximum female and male body size Across a species distribution growth rates influencing life history and demography can vary considerably due to an interaction between genetic and environmental regulation Several issues have hindered understanding of the causes of individual growth rate variation in animals, and particularly ectotherms that experience indeterminant growth. First, most growth rate models do not accommodate potential polyphasic growth Varanus komodoensis; 300 cm snout-vent-length and 87 kg body mass). Data were collected from marked individuals from 2002–2010, with the aim to evaluate individual, sex-specific and spatial patterns in somatic growth. It was assumed that several energy allocation trade-offs may alter investment in growth in Komodo dragons during life-history changes including distinct transitions in habitat use, sex related growth differences pre- and post-maturation, and aging, that may occur in long-lived apex predators such as this, which have very low mortality due to predation . Each of these phenomena could result in polyphasic growth trajectories involving growth “lags” and “spurts” across ontogeny. As this sort of pattern may not be possible to model with the use of standard growth models such as the von Bertalanffy growth function  in this species  was addressed. Such pronounced SSD must entail different tactics and requirements for energy acquisition and investment The final aim considered spatial variation; looking at differences in ecological, demographic and genetic factors on the islands that may account for differences in growth patterns among sites. Dragon body size varies dramatically between islands, with the presence of dwarf and giant forms in part suggesting spatial variation in growth. For Komodo dragons potential variation in growth could be mediated by differences in prey availability Rusa timorensis), feral pig (Sus scrofa) and water buffalo  The Komodo dragon is endemic to five islands in eastern Indonesia, with four island populations in Komodo National Park and several fragmented populations on Flores 2 Komodo Island); Loh Baru (Lba), Loh Buaya (Lbu), Loh Dasami (Lda) and Loh Tongker (Lto) ; and one site on each of the islands of Gili Motang  and Nusa Kode . Only two of 1062 marked Komodo dragons moved between any of the sites during the course of the study  and hence the ten sites are considered discrete closed populations.To evaluate growth patterns in Komodo dragons, a capture-mark-recapture (CMR) study was undertaken at ten sites on four islands in Komodo National Park , eastern Indonesia , during Dragons were captured in a trapping grid at each site using aluminium box traps and noose poles, and were uniquely marked with passive integrated transponder (PIT) tags . Further details of the capture program are outlined elsewhere Absolute growth rates were calculated from growth records for each individual sampled using snout-vent-length as the dependent variable, and included negative and zero growth rates because animals can shrink in size due to senescence or in response to extreme resource limitation A two-stage statistical modelling approach was used to model somatic growth Size-specific growth rates were modelled as a function of snout-vent-length using a GAMM t- and F-ratio tests The mgcv package Size frequency distributions were constructed for all male and female lizards using the SVL obtained at the first capture event. Using sex-specific age-at-size equations we predicted the mean age for each individual at first capture. To evaluate if male and female size and predicted age distributions differed significantly we performed Kolmogorov-Smirnov tests in program R is) (see methods below for determination of site estimates for each covariate). The four candidate models, including a null model, were fitted again using a GAMM approach in the mgcv package w), considering the strength of evidence that the candidate model is the best model for the data Four competing models were conceived involving three putative covariates; population density, prey availability, and level of inbreeding  Faecal counts were conducted annually from 2003–2010 (i.e. late dry season) at each site, with faeces of Timor deer and water buffalo counted within circular plots on 150 m transects in each site. Hand-held GPS  units were used to locate transect line starting points, and ungulate faeces were counted within 30 sample plots placed at 5 m intervals along each transect. Between 20 and 41 transects were randomly positioned and orientated at each site, providing a total of 308 transects with a total length of 45.50 km. Faecal counts of both species show positive relationships with actual density estimates derived from distance sampling V. komodoensis genetic microsatellite data sampled from eight sites is) were calculated using Genetix 4.01 is values to a frequency distribution of fixation indices obtained after 10,000 permutations of alleles.Estimates of site-specific inbreeding coefficients were obtained predominantly from previously analysed This Research was authorized under successive collaborative research memorandums of understanding (MOU), first (2002–2007) between Zoological Society of San Diego, The Nature Conservancy and the Indonesian Department of Forest Protection and Nature Conservation (PHKA), and second (2008–2015) under MOUs between the Komodo Survival Program and PHKA. Animal experimental ethics committee approval was obtained from the University of Melbourne (under Permit 0911162.1).Growth measurements were obtained from 400 individually marked Komodo dragons refer to  capturedThe estimated size-specific growth function for Komodo dragons was polyphasic with highest growth rate at the juvenile stage , and incComparing the estimated size-specific growth curves of both sexes  however,The estimated size-at-age growth curves indicate that males and females are of similar size until ∼7 years , after wThe estimated age-specific growth function  indicateMale and female lizards exhibited significantly different frequency distributions in body size with males exhibiting substantially larger body sizes than females  exhibited: distinct sex-based differences in growth trajectories, suggesting altered energy allocation tactics with different life-stages; and spatial variability in growth rates, due to population density-dependent processes.Somatic growth dynamics of individuals and populations can have profound consequences for fitness, life-history and demography In species with indeterminate growth, where females have higher energy costs for reproduction, they must allocate nearly all energy investment from growth to reproduction after maturity ; whereas males may typically invest considerably less to ensure reproductive success increases with body size The largest females tend to be in poor body condition  due to extended periods of fasting whilst nest-guarding. Males in comparison appear to live longer presumably aided by lower reproductive costs and an absence of predation conferring higher male survival. A consequence of these sex related differences appears to be large differences in the age of maturity and also maximum longevity estimates . Female Significant variation in mean growth rates was evident among sites and islands . SupportWith size-specific growth variation among sites and islands, it would be worthwhile to detect specific habitats with distinct environmental or genetic differences in dragon growth trajectories that underpin life-history traits such as maximum body size, longevity and vital rates. Vast geographic variation in phenotypic traits are recognised in various intra- and inter-specific studies of sea turtles Our study shows that across ontogeny somatic growth of Komodo dragons is polyphasic in males, yet not in females, as a consequence of differences in energy allocation tactics regarding the onset of reproduction, which also result in an extreme contrast in longevity between sexes. This vast difference in life-span likely has a significant effect on population sex ratios, which in turn could be increasing sexual size dimorphism (SSD) within this species. In future it would be interesting to compare how SSD may vary between populations in regards to demographics and environmental quality Figure S1Mean growth year index as a predictor for growth rate in Komodo dragons. Year index was one covariate in the fitted generalized additive mixed model (GAMM). Dotted lines represent the 95% confidence interval for the fitted values.(TIF)Click here for additional data file.Figure S2Spatial variation in growth rates in Komodo dragons. Mean growth rates (cm SVL/yr) for each site indicate spatial variation in growth among sites and islands. Error bars are standard errors of site mean growth rates.(TIF)Click here for additional data file.Methods S1(DOCX)Click here for additional data file.Methods S2Methods & Results.(DOCX)Click here for additional data file.Table S1(DOCX)Click here for additional data file.Table S2(DOCX)Click here for additional data file.


Dear Editor,The study named “The role of myocardial perfusion gated SPECT study in women with coronary artery disease” reveals some current and important issues in diagnosing myocardial ischemia in women . BecauseOn the other hand the evaluation criterion for diagnosing ischemia seems controversial in the study. The question is that: “Do the left ventricular (LV) wall motion abnormality and decreased systolic thickening at the segment with reduced perfusion in gated SPECT study necessarily show true ischemia as it is expressed in the study?This approach considers that wall motion abnormality and perfusion defects occur exactly at the same time in CAD stages, but as it is known, wall motion abnormalities follow quite a bit later after the perfusion defects appear. Myocardial hypoperfusion is first seen in ischemic cascade, and followed by decrease in metabolic activity, relaxation impairment (diastolic dysfunction), reduction in contractility (systolic dysfunction), global LV dysfunction, ECG changes and angina pectoris, respectively . So, theMore importantly, because the imaging begins 30-60 minutes after the radiopharmaceutical injection, stress–gated images with Tc-99m MIBI show stress perfusion at the time of injection, but LV function  at the time of acquisition. Snapper et al. demonstrated that the combination of a perfusion defect with normal wall motion and thickening in the same segment on stress-gated MIBI images was associated with a high positive predictive value (96%) for detecting ischemia. They concluded that the diagnosis of ischemia could be made confidently in patients with normal wall thickening in a segment with perfusion defect. The lack of wall thickening was found less specific, because only 40% of the cases with perfusion defects and wall motion abnormalities showed reversibility on rest perfusion imaging . Thus, tFinally, the criterion mentioned in the study could be entirely true and supporting if all the subjects had been shown to have reversible defect (ischemia), but some patients are reported to have myocardial infarction. So the criterion might include myocardial scar in patients with fix defect (particularly when the presence of metabolic activation on the defect related with hibernation was excluded). In the routine evaluation of gated SPECT studies, wall motion abnormality and/or systolic thickening at the same segment with fixed defect primarily indicates myocardial scar. So, the inclusion criterion that the authors used in the study constitutes a great limitation for drawing true conclusions about ischemia and/or infarct without making some comparisons between post-stress and rest end-systolic and end-diastolic volumes or measuring LVEF differences together with the wall motion abnormalities.The evaluation criteria to differentiate ischemia, infarct and soft tissue attenuation artifacts in stress-rest gated SPECT imaging may be simply summarized as below:1. Normal perfusion with no wall motion abnormality or TID: Normal2. Normal perfusion with wall motion abnormality or TID: Stunning (and/or multivessel disease?).3. Reversible ischemia with no wall motion abnormality, TID or reduced LVEF in stress-gated images: Ischemia.4. Reversible ischemia with wall motion abnormality, TID or reduced LVEF in stress-gated images: Severe ischemia with increased cardiac event rate.5. Fixed defect with normal wall motion or wall thickening: Soft tissue attenuation artifact.6. Fixed defect with wall motion or wall thickening abnormality: Myocardial scar.


Cryopreservation and transplantation of ovarian tissue is one option for re-establishing ovarian function, but optimal conditions for graft sustainment and follicular survival are still considered experimental. The present study aims to analyze the effect of FSH treatment on the resting follicle pool in fresh and cryopreserved primate ovarian tissues following xenografting.Ovarian tissues from adult marmosets were grafted freshly or following cryopreservation to ovarectomized nude mice treated with FSH 25 IU twice daily post transplantation or left untreated as controls. Grafts were retrieved 2 or 4 weeks after transplantation to evaluate the number and morphological appearance of follicles.Early start of FSH treatment within 1 week following transplantation partly prevents primordial follicle loss in fresh and frozen-thawed tissues, whereas after a 3 weeks time interval this effect is present only in fresh tissues. A similar positive effect of early, but not later FSH treatment on primary follicles is seen in fresh tissues compared to only marginal effects in frozen-thawed tissues. The percentage of morphologically normal follicles is generally increased in FSH treated tissues, whereas the percentage of primary follicles over all primordial and primary follicles is increased by FSH only in freshly-grafted tissues.FSH treatment alleviates depletion of the resting follicle pool and promotes normal follicular morphology both in freshly and frozen-thawed grafted tissues. In previously cryopreserved tissues, applying to most of the tissues intended for clinical use in fertility preservation attempts, its positive effect on primordial follicle numbers and potential graft sustainment is dependent on an early start of treatment within one week of transplantation. Increasing cancer survival rates and awareness of fertility as a quality of life issue in long-term cancer survivors have fuelled the growing demand for fertility preservation in recent years . In femain vitro study has confirmed equal effectiveness of human recombinant FSH compared to marmoset FSH on the marmoset FSH receptor . Grafting duration has no influence on mean primordial follicle numbers  compared to untreated tissues  but not at 4 weeks  and frozen-thawed (29.9+/−2.9%) ovarian tissues. In freshly grafted tissues, these proportions decrease significantly after transplantation and then remain stable over time , but not FSH-treated tissues (p=0.411; Figure The size of the resting follicle pool determines the ovarian reserve of an individual and correlates with graft sustainment after restoration of fertility by ovarian tissue cryopreservation and transplantation. Assessment of ovarian reserve following ovarian tissue transplantation is challenging when using established markers and indicates high variability between patients . Besidesnd week of transplantation. In freshly but not in frozen-thawed grafted tissues, a positive effect of FSH treatment on primordial follicle number was also observed if the start of treatment was postponed to 3 weeks post transplantation. As we have previously documented in the marmoset [in vitro co-culture system had previously been described for other non-human primates [Enhanced follicular depletion through FSH effects during ovarian stimulation has anecdotally been reported in the human , but conmarmoset , and othmarmoset ,33,34, cprimates . In humaprimates , thus enprimates , and forprimates , suggestWhen started after a prolonged grafting period of frozen tissues, FSH negatively influences primordial follicle numbers that are already low and most likely at this point reflect manifestation of preceding tissue damages due to cryopreservation. Since primary follicle numbers are increased in these tissues, the reduction in the primordial follicle pool may additionally mirror the beginning of follicular recruitment becoming apparent in already low primordial follicle numbers. However in fresh tissues, baseline values of follicle numbers are higher also after a prolonged grafting period, and FSH treatment following early or at a later point in time after transplantation prevents primordial follicle loss. This is in agreement with viability studies on fresh and frozen ovarian tissue resulting in significantly reduced viability when applying a slow-freezing protocol .per se on follicle pool dynamics and primary follicle activation in adult and prepubertal ovarian tissues [Proportional analysis of primary and primordial follicle number can indicate initiation of follicular growth, and for xenografted human ovarian tissue the influence of cryopreservation  and timi tissues . This co tissues . As the FSH treatment alleviates depletion of the resting follicle pool and promotes normal follicular morphology both in freshly and frozen-thawed grafted tissues. In previously cryopreserved tissues, applying to most of the tissues intended for clinical use in fertility preservation attempts, its positive effect on primordial follicle numbers and potential graft sustainment is dependent on an early start of treatment within one week of transplantation. Whereas in fresh tissues FSH treatment following xenografting results in a higher percentage of primary over all un-advanced follicles, this appears to be largely independent of FSH in frozen-thawed tissues.All authors declare that they have no competing interests.VVS and BS designed the study, analyzed and interpreted the results and drafted the manuscript. RC performed the experiments and analyzed the results. EN and LK participated in the study design, helped in the acquisition of funding and critically reviewed the manuscript. RO helped to perform statistical analysis and with the drafting of the manuscript. All authors read and approved the final manuscript.


Duchenne muscular dystrophy (DMD) is a severe, progressive muscle-wasting disorder, while Becker muscular dystrophy (BMD) is milder muscle disease . Both arThe exon skipping approach uses antisense oligonucleotides (AONs) to induce skipping of targeted exons during pre-mRNA splicing, with the aim of reading frame restoration, converting of the severe DMD into the milder BMD phenotype . This apAfter promising results in cultured cells and animal models where AON treatment allowed in dystrophin restoration (reviewed in ), a firsTowards systemic application, studies in animal models revealed that dystrophic muscles facilitated uptake of 2OMePS AONs and that subcutaneous delivery was feasible . In a suIn parallel, preclinical studies to further optimise treatment regimens are in progress as well as clinical trials for additional exons for exon 44 skipping . Trials are planned for exon 45 and 53 skipping .The mutation specificity of the approach poses challenges to drug development regulations. A concerted effort of academic researchers, industry, regulators and patients is needed to adapt regulations to enable application of these personalised medicine approaches to rare diseases.


Olfactory dysfunction develops in many neurodegenerative diseases, and is an early feature of the most common neurodegenerative disorder, Alzheimer's disease (AD).Here we compared olfactory function prospectively in cohorts of patents with PCA and typical AD (tAD). Neuroanatomical associations of odour identification were assessed using voxel-based morphometry (VBM). We hypothesised that PCA would be associated with olfactory impairment qualitatively similar to tAD, but less severe ; and that deficits of odour identification in both syndromes correlate with grey matter loss in anteromedial temporal and inferior frontal lobes.6–10Fifteen patients fulfilling consensus criteria for PCA,supplementary table S1). Further details about the behavioural assessments are in online supplementary material.All subjects had a comprehensive general neuropsychological assessment which corroborated the clinical impression in both disease groups (see online supplementary table S2) before identifying it; and target-foil choices were name-picture combinations rather than odour names alone, to maximise available response cues. Group differences were assessed using analysis of variance (ANOVA) or χ2 tests (Stata V.12.1), adjusting for cognitive severity, verbal processing measures, age and gender.Olfactory processing was assessed using the 40-item, four-alternative-forced-choice University of Pennsylvania Smell Identification Test (UPSIT: British version).http://www.fil.ion.ucl.ac.uk/spm) following previously described procedures . Linear regression was used to examine voxel-wise associations between regional grey matter volume and odour identification performance (age-normed and gender-normed percentile score) across the combined patient cohort, within the PCA subgroup, and between syndromic subgroups, incorporating syndromic group, mini-mental state examination (MMSE) score and total intracranial volume as covariates. Statistical parametric maps were assessed thresholded over the whole brain volume and after multiple-comparisons correction over small volumes of interest  specified in our prior anatomical hypotheses.Twelve patients with PCA and eight patients with tAD had T1-weighted volumetric magnetic resonance (MR) brain images acquired on a 3.0T Siemens Trio scanner. VBM was performed using SPM8 . Based on published UPSIT norms,supplementary table S1). An error analysis of individual odour items in the identification test revealed a qualitatively similar profile of errors across all groups (see online supplementary figure S2).For both patient groups, mean odour categorisation and identification raw scores were significantly lower than the HC group . Across the combined patient cohort, performance on the odour identification task was positively associated with regional grey matter volume in right entorhinal cortex and parahippocampal gyrus . At a more lenient threshold (p<0.001 uncorrected over the whole brain volume), additional associations were present in more distributed, predominantly right-sided cerebral areas, including hippocampus, posterior inferior temporal gyrus/sulcus, temporo-parieto-occipital junction and premotor cortex (see online supplementary table S3). Similar grey matter associations of odour identification performance were identified for the PCA subgroup alone . Direct comparison between the PCA and tAD subgroups revealed no significant between-group differences in regional grey matter correlations of olfactory performance.Statistical parametric maps of significant regional grey matter associations of odour identification performance are displayed in Here we have demonstrated deficits of odour identification and categorisation in patients with PCA relative to HCs. A similar proportion around 30%) of patients with PCA and tAD in this study had an absolute deficit of odour identification referenced to published age and gender norms and taking account of associated cognitive impairment. Olfactory impairment was similar quantitatively and qualitatively in the PCA and tAD groups. To the extent that PCA manifests underlying AD, the findings imply that olfactory impairment is a hallmark of AD pathology. It is noteworthy that only a minority of patients in both phenotypical groups here reported olfactory symptoms, suggesting that in many cases olfactory impairment is ‘subclinical’. Mean corrected odour identification scores were higher than categorisation scores in the HC and PCA groups: this unexpected finding might hold clues to the cognitive organisation of olfactory knowledge or the cognitive strategies engaged by these tests, and would warrant further study in larger populations. Odour identification tasks tend to be cognitively demanding and therefore potentially susceptible to executive and attentional deficits that accompany AD.0% of pat13The deficit of odour identification identified here was associated with regional grey matter volume in a cerebral network focussed on the right anteromedial temporal lobe. The most robust neuroanatomical associations occurred in parahippocampal gyrus and entorhinal cortex: areas linked to odour identification in healthy human subjects.This study has several limitations that suggest directions for future work. We did not assess perceptual encoding of odours: it will be important to compare associative and perceptual olfactory functions directly, to assess the extent to which these different factors contribute to olfactory impairment in AD. The patient groups studied here were relatively small: there is a need to extend the work to larger patient cohorts spanning other AD phenotypes  and in direct comparison with other neurodegenerative pathologies.


After publication of our work , we notiThe authors declare no competing interests.JL and DM designed research; JL carried out phylogenetic analyses, and JL and DM wrote the manuscript. Both authors read and approved the final version.