Intracardiac blood flow measurements using 4D Phase Contrast CMR (4D PC-CMR) may contain useful information about cardiac pumping. Lagrangian Coherent Structures (LCS) is a new, operator-independent method which can simplify analysis by partitioning the flow into regions with different origins and destinations. Since LCS are operator-independent, they may be used to define quantitative indices of intracardiac blood flow. The partitioning has not previously been validated against particle tracing in 4D PC-CMR blood flow.To investigate whether Lagrangian Coherent Structures computed from 4D Phase Contrast CMR of the human heart partitions diastolic inflow blood from blood already in the left ventricle.Eight healthy volunteers  underwent 4D PC-CMR flow measurements of the whole heart. Three-dimensional LCS surfaces were computed and automatically delineated in the left ventricle during diastole. The only parameter adjustable by the operator was the start of diastole. Particle tracing was performed to study the blood flowing into the ventricle during diastole and the blood already present in the ventricle from the previous heartbeat. LCS and particle traces were compared visually using the software Ensight .In all eight subjects, a distinct LCS was found in the left ventricle during diastole. The LCS separated the inflow blood particle traces and particle traces of blood already in the ventricle in all eight subjects. The Figure shows the typical appearance of the LCS and particle traces in one healthy volunteer.Lagrangian Coherent Structures can separate 4D Phase Contrast CMR data into regions of blood with different origins. Since Lagrangian Coherent Structures are operator-independent, they have the potential to be used to define quantitative indices of intracardiac blood flow.


This percentage decreases in faster runners, where the incidence of midfoot and forefoot strikers (FFS) increases . It is pet al.  found ru-1 while muscle fibre length change of the medial and lateral gastrocnemius and soleus were recorded using ultrasound and muscle activation via surface electromyography. Individual contribution of the triceps surae muscles to Achilles tendon force was determined using a Hill-type model based on muscle activation, the muscles force-length-velocity relationship , maximum isometric muscle force and pennation angle. Achilles tendon and triceps surae individual muscle forces were recorded while runners performed their natural (fore or rearfoot) and then converted to their unnatural strike.Natural FFS (n=9) and RFS (n=9) distance runners ran on a treadmill at 3msResults from one FFS participant are presented below Figure  &2, addiThis research provides insight into the role of the Achilles tendon during FFS running and sheds light on its’ contribution to reducing energy cost. It also reveals the altered demand on the triceps surae muscles which may have implications for technique recommendations and training requirements.


Synaptic dysfunction is widely thought to be an important pathogenic event in Alzheimer’s disease (AD). Presenilins, which harbor large numbers of mutations for familial AD and frontotemporal dementia (FTD), are important for neurotransmitter release and synaptic plasticity as well as memory and age-related neuronal survival.2+ imaging coupled with electrophysiological and molecular analyses using both acute hippocampal slices of unique presenilin conditional mutant mice and cultured hippocampal neurons, in which presenilins are acutely inactivated with a lentivirus expressing Cre recombinase.Our recent report showed that presenilins regulate synaptic function by modulating ryanodine receptor-mediated calcium release from the ER. To determine how presenilins regulate intracellular calcium signaling in neurons, we performed Capresenilin-1 knockin mice, in which either an AD- (L435F) or FTD (G183V) -causing mutation is introduced into the respective presenilin-1 endogenous genomic locus, to investigate the mechanisms by which presenilin mutations cause AD or FTD. The analysis of these mutant mice will also be presented.Our results reveal a selective interaction between presenilins and ryanodine receptors in the regulation of calcium homeostasis and synaptic function, and suggest that disruption of calcium homeostasis may be an early pathogenic event leading to synaptic dysfunction in AD. We also generated two


A genetic predisposition to cluster headache (CH) has long been debated. Genetic epidemiological studies have reported an increased risk of CH in relatives of CH sufferers. The familial clustering supports a model of autosomal dominant inheritance with reduced penetrance. Some candidate gene studies have been performed, detecting associations with HCRTR2 and ADH4, but these have been limited by small sample size. We have established a large cohort of CH families in which we have previously reported a genome-wide linkage scan, isolating a number of putative linkage loci. DespiteTo further delineate the genetic architecture underlying CH, we have used an exome sequencing strategy in a subset of Northern European families.Exome target enrichment and paired-end sequencing were performed for ten probands. Annotated variants were filtered to exclude known polymorphisms, leaving a total of 1711 novel variants. Exome data from related affected subjects were examined to limit the analysis to variants segregating with the CH phenotype. Segregation analysis by Sanger sequencing in all family members reduced the candidate list to a total of 45 genes (range: 1-13 genes per pedigree). These genes are now being screened in our extended cohort to provide further insight into the role of each gene in CH pathogenesis.Whilst exome sequencing for rare monogenic disorders is now well-established, approaches to detect pathogenic variation with complexities such as locus heterogeneity and incomplete penetrance remain challenging. We have combined exome and Sanger sequencing to isolate novel coding variation segregating across CH families, highlighting the need for large homogeneous cohorts to elucidate the molecular genetic basis of CH. The significance of these variants in CH pathogenesis remains to be determined; however these results provide further evidence for a potential genetic predisposition to this debilitating disorder.


A single center, cluster randomized controlled effectiveness trial was conducted to compare two novel hand hygiene (HH) promotional strategies. Sixty six wards were randomized to standard HH promotion (control), standard promotion plus HH performance feedback (FB), or FB plus active patient participation (PP). While FB seemed to be well accepted by HCWs, the introduction of PP proved to be more challenging.Here we focus on PP and report the results of a qualitative inquiry, designed to investigate the range of success with its implementation.A ward-level case study was conducted through 3 focus groups with ward staff and 6 interviews with ward head nurses. Participants were selected following an extremes sampling strategy regarding adoption strength. The hospital’s infection control nursing staff participated in additional focus group. All sessions were audiotaped and transcribed verbatim. Analysis was deductive (using a framework of themes that had been previously established) and inductive (grounded theory).Four main and 12 sub-themes (in parenthesis) emerged: Context ; Risk knowledge (nil); Interaction ; Implementation process . The action of confronting another person about a failure became less emotionally menacing through positive experiences, innovative means of communication, and institutionalization of the project. Individual leadership engagement had a major impact on implementation success. Exclusion from intervention arms motivated control wards to improve HH performance independently.This qualitative research permitted to explain the quantitative study findings and to explore the evolutionary and complex implementation process of PP. While PP remains challenging, it is our hope that the findings of this study may facilitate future patient participation projects.None declared


Juvenile Fibromyalgia (JFM) is characterized by chronic widespread musculoskeletal pain and approximately 40% of children and adolescents with JFM also suffer from benign joint hypermobility (HM). It is not currently known if the presence of HM affects the pain experience of adolescents with JFM. The objective of this study was to examine whether there were any differences in self-reported pain intensity and physiologic pain sensitivity between JFM patients with and without joint HM.One hundred thirty-one adolescent patients with JFM recruited from four pediatric rheumatology clinics completed a daily visual analogue scale (VAS) pain rating for one week and underwent a standardized 18-count tender point (TP) dolorimeter assessment. Medical records were reviewed for the presence of joint HM. Average pain VAS ratings, tender point count and tender point sensitivity were compared between JFM patients with and without hypermobility (HM+ and HM-).Nearly half (48%) the sample of JFM patients were found to be HM+. HM+ and HM- patients did not differ in their self-reported pain intensity. However, HM + patients had significantly greater pain sensitivity, with lower TP thresholds (p = 0.002) and a greater number of painful TPs (p = 0.003) compared to HM- patients.The presence of HM among adolescent patients with JFM appears to be associated with enhanced physiologic pain sensitivity, but not self-report of clinical pain. Further examination of the mechanisms for increased pain sensitivity associated with HM, especially in adolescents with widespread pain conditions such as JFM is warranted. Juvenile fibromyalgia (JFM) is a chronic condition of widespread musculoskeletal pain and fatigue in children and adolescents. Prevalence estimates for JFM range from 1-6% of the pediatric population . While tBenign joint hypermobility (HM) is a relatively more common condition than JFM, with prevalence rates in children and adolescents estimated to be up to 30% . ChildreThe underlying mechanisms for pain hypersensitivity in FM have been extensively studied while the etiology of pain in HM has received little attention. It has been well documented that persons with FM have an overall lower threshold for pain as demonstrated by increased responsiveness and hypersensitivity to pain  in the fAs part of the screening for a larger clinical trial examining the effectiveness of cognitive-behavioral therapy for JFM, we assessed baseline pain intensity and tender point sensitivity (tender point count and tender point threshold) of over 100 adolescents with JFM . For theParticipants were 131 adolescents  with JFM between the ages of 11 and 18 years (mean age = 15.1 years) who were initially screened for the parent clinical trial. Participants were recruited from four pediatric rheumatology clinics  in Ohio and Kentucky, with each site having Institutional Review Board approval. All participants met Yunus and Masi criteria .


Purulent mediastinitis is a possible serious complication after mediastinal surgery. We report the case of a localized sternal plasmocytoma treated by sternectomy and prosthetic repair, who needed a second surgery for a fistulizing mediastinitis. Five months earlier, in another Hospital, the patient underwent sternal resection and reconstruction with a “sandwich” prosthesis (Methyl-methacrylate and Marlex mesh). Suppurative mediastinitis occurred and septic shock resolution was observed after the spontaneous opening of a mediastinal cutaneous fistula. After referring to our Unit the patient underwent extensive local and systemic preparation and nutritional support; the infected prosthesis was then removed and the gap filled by a laparoscopically-prepared omental flap. Adequate preoperative management, removal of any infected material and minimally invasive omental flap transposition allowed the successful treatment of this life-threatening condition. Purulent mediastinitis secondary to chronic prosthetic infection after sternectomy is a very difficult problem to resolve. The goals of surgical treatment are fast removal of the infected prosthetic material, mediastinal debridement and mediastinal filling with well vascularized tissues. Surgical trauma should be as mild as possible for the usually weak patient’s general conditions ,2.We describe the case of a 73-year-old caucasian male with purulent mediastinitis and prosthetic infection after subtotal sternectomy.Five months earlier, the patient underwent surgery for solitary plasmocytoma in another hospital; chest wall reconstruction was achieved by a Marlex methyl-methacrylate sandwich prosthesis. Two days after surgery, the wound was re-explored to control bleeding from the operative site.At the admittance to our hospital the patient was in a bad clinical condition, with intermittent fever, dyspnea and anterior chest pain. His comorbidities included chronic C virus hepatitis, monoclonal gammopathy and multinodular goiter. Blood sampling revealed an increase in white blood cells, erythrocyte sedimentation rate (ESR) and C-reactive Protein (CRP). Local examination showed an erythematous, swollen, indurated sternal wound with a chronic, purulent fistula opening at the lower third of the surgical incision.A computed tomography (CT) scan showed semi-solid inflammatory tissue with multiple air levels involving the mediastinal surface of the sternal prosthesis and the pericardium Figure A-B. MicrUnder general anesthesia with a single lumen oro-tracheal intubation, we laparoscopically prepared an omental flap, transposed at the thoracic level through a minimal diaphragmatic anterior incision Figure A. The tePatient was discharged on postoperative day 10 and received intravenous antibiotic therapy for another 2 weeks. Drains were removed on postoperative day 4; ESR and CRP decreased towards baseline values within 47 days. The wound healed well without significant flail chest (Figure A rigid plaque as methyl-methacrylate in combination with two layers of Marlex mesh has been largely employed to repair large defects after sternectomy. The possible infection of this type of prosthesis is already known  and has Therapeutic alternatives not entailing the removal of the infected prosthesis are to be considered unreliable. In fact sepsis resolution can be achieved only after the thorough removal of all the infected autologous and prosthetic tissues .•Removal of an infected sternal prosthesis is the key for the successful treatment.•Intrathoracic omental flap transposition is preferable to muscle flap but entails an heavier surgical trauma.•Laparoscopically-prepared omental allows to take advantage from the favorable properties of the omentum, without further significant surgical trauma.“Written informed consent was obtained from the patient 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.”ESR: Erythrocyte sedimentation rate;CRP: C-reactive protein;CT: Computed tomographyAll authors declare that they have no competing interest.VT, SC and FP wrote the article, JV and SC collected the clinical information, JV selected the images; VT and SC analyzed the English Literature. FP drafted the final manuscript. All authors approved the final manuscript to be published.


Plasmodium (P.) synthetic lethal (SL) gene pair has antimalarial drug resistance (ADR), a drug targeting the other gene of the SL pair can be used as an effective antimalarial drug for treating drug-resistant strains of malaria diseases. The approach introduced in this study is based on bioinformatic database integration. It employs a promising concept of synthetic lethality and has practical applicability in identifying gene targets to discover potential future drugs.If one gene of the P. genes were searched to find new drugs for clinical malaria treatment from Yeast Fitness DB [Potential antimalarial drug target genes were identified by integrating whole biological information from many databases, including BioGRID , KEGG SStness DB .P. species  was established to identify SL genes that are possible drug targets. Information on SL gene pairs with ADR genes and their first neighbors from the yeast SL gene was inferred to search for pertinent antimalarial drug targets. We suggested that specific antimalarial drug candidates can be inferred by searching drugs that cause a fitness defect in the yeast SL gene.A simple computational tool to analyze inferred SL genes of P. Malaria parasites can be killed by mutating or blocking the SL partner of ADR genes. This concept is useful in selecting candidates as drug targets in antimalarial therapy. The methodology also provides not only drug target gene candidates for further experimental validation, but also information on new usage for already-described drugs. Drug candidates for targeting the suggested genes significantly benefit experimental validation in antimalarial drug discovery.We suggest a new concept of drug-resistant malaria therapy in this study. The alternative antimalarial drug therapy consists of data integration and inference through the homology analysis of yeast-human-


Dear SirThis is in reference to our article published before [1]. On 1 year follow up, the patient had local recurrence measuring 8 cm X 5 cm infero-laterally to the previous scar. MRI of the local region showed a large well defined heterogeneously enhancing soft tissue mass in subcutaneous layers involving ilio-tabial tract with areas of deep multiple recurrences . There was no evidence of metastasis. Local excision was done. Histopathology was identical as before- the desmoid tumor. The surgical margins were free; the mass was surrounded with fibrocollagenous tissue. Keeping in view the unresectable deep recurrences, he was administered radiotherapy locally. MRI done after 6 weeks of radiotherapy did not show any residual disease. The child is under close follow up.Even after multiple recurrences, successful salvage is achievable, particularly when high-dose focal radiotherapy is incorporated [2]. It would also be imperative to mention role of intra-operative electron radiotherapy (IOERT), followed by moderate doses of external beam radiotherapy (EBRT) after organ-sparing surgery, in patients with primary or recurrent aggressive fibromatosis. Introduction of IOERT into a multimodal treatment approach in patients with aggressive fibromatosis is feasible with low toxicity and is known to yield good local control rates even in patients with microscopical or gross residual disease [3]. Such facility is however unavailable in our setup.In our case, the hip joint was not involved. A case of aggressive pediatric hip fibromatosis with severe joint destruction has been reported recently [4], and we would like to prepare ourselves for such an eventuality. Source of Support: NilConflict of Interest: None declared


The aim of our study was to test the robustness and efficiency of maximum likelihood with respect to different long branch effects on multiple-taxon trees. We simulated data of different alignment lengths under two different 11-taxon trees and a broad range of different branch length conditions. The data were analyzed with the true model parameters as well as with estimated and incorrect assumptions about among-site rate variation. If length differences between connected branches strongly increase, tree inference with the correct likelihood model assumptions can fail. We found that incorporating invariant sites together with  Maximum likelihood (ML) tree inference has been shown to be statistically consistent for binary trees with finite branch lengths under correct model and model parameter assumptions as sequence length increases to infinity The ML method is certainly more robust and more efficient than other methods It is well known that if among-site rate variation (ASRV) is ignored in tree reconstruction, the ML approach underestimates substitution rates, and these estimates become progressively worse with increasing evolutionary distances Early studies argue that combining both models  predominates tree inference even under correct model assumptions (settings . In geneTopology B was designed to test for class I effects (symplesiomorphy effect). The major difference to topology A is that the evolving sequence passes through two long branches, while in topology A the two long branches are parallel. Even if the correct proportion of invariant sites  correct as well as estimated model parameters . This has also been observed in previous studies, e.g. The inclusion of a mixed-distribution model , is not a particular problem of mixed-distribution models. If no invariant sites are estimated in the reconstruction, this model deficiency is partially compensated by a lower estimated value of the Our results show that the risk of obtaining a wrong topology using ML is dependent on the arrangement of the edges (corresponding to which LBA classes the tree is susceptible to). Although our results depend on simulated nucleotid data it can be expected that amino acid sequences are also prone to long branch effects if branch lengths combinations of RB) are kept constant (We designed two sets of data simulations under different topologies  to detecn length . This seranch  . This trTrees were inferred with the Jukes-Cantor (JC) model under different parameter settings using PhyML-3.0-linux64 Wrong topologies were classified into LBA class I, II and III effects . Wrong tFigure S1ML reconstruction success of simulated paramter and branch length settings.(TIFF)Click here for additional data file.Figure S2ML values of reconstructed topologies.(TIFF)Click here for additional data file.Figure S3ML parameter estimates of reconstructed topologies.(TIFF)Click here for additional data file.


The integrate-and-fire (IAF) neuron model is a Time Encoding Machine (TEM), which maps analog signals into a sequence of strictly increasing time events. There are a number of reconstruction algorithms that ensure perfect recovery of bandlimited input signals from spike trains . For sig


The cGMP-dependent protein kinase (PKG) has two tandem cyclic nucleotide binding (CNB) domains which act as the primary intracellular receptor for cGMP ,2. PKG e78-355 structure is free of cGMP and presents the protein in an elongated conformation. A surprising dimeric arrangement between PKG78-355 protomers is orchestrated via hydrophobic contacts between a novel helical element C-terminal to the second cGMP binding site (the switch helix) and the opposite CNB domain B , suggesting the additional importance of the switch helix in mediating cGMP-specific conformational changes inherent to the regulatory domain.We recently solved a crystal structure of the two cGMP binding sites from PKG Iα in order to highlight the atomic details of the regulatory domain. This PKGB Figure . Small aOverall, these studies provide the first atomic resolution model of tandem cGMP binding domains and expand our understanding of the allosteric mechanisms surrounding PKG activation.


Interspecific divergence along a benthic to pelagic habitat axis is ubiquitous in freshwater fishes inhabiting lentic environments. In this study, we examined the influence of this habitat axis on the macroevolution of a diverse, lotic radiation using mtDNA and nDNA phylogenies for eastern North America’s most species-rich freshwater fish clade, the open posterior myodome (OPM) cyprinids. We used ancestral state reconstruction to identify the earliest benthic to pelagic transition in this group and generated fossil-calibrated estimates of when this shift occurred. This transition could have represented evolution into a novel adaptive zone, and therefore, we tested for a period of accelerated lineage accumulation after this historical habitat shift.Ancestral state reconstructions inferred a similar and concordant region of our mtDNA and nDNA based gene trees as representing the shift from benthic to pelagic habitats in the OPM clade. Two independent tests conducted on each gene tree suggested an increased diversification rate after this inferred habitat transition. Furthermore, lineage through time analyses indicated rapid early cladogenesis in the clade arising after the benthic to pelagic shift.A burst of diversification followed the earliest benthic to pelagic transition during the radiation of OPM cyprinids in eastern North America. As such, the benthic/pelagic habitat axis has likely influenced the generation of biodiversity across disparate freshwater ecosystems. Freshwater fish are frequently thought to diversify along a benthic (bottom) to pelagic (mid-water) habitat axis-3. HowevClinostomus elongatus and C. funduloides) and five endemic to western North America  form the sister group to a much larger group of species that is primarily confined to eastern North America.All trees are deposited in TreeBASE  Cytb and (B) Rag1 MCC gene trees for the primarily eastern North American OPM cyprinid radiation. Numbers at nodes represent posterior probability values (pp). Asterisks denote 100% pp. The ‘transition nodes’ indicated by black arrows correspond to those in Figure Click here for fileDensity plots for lineage accumulation statistics and ΔAIC values from 1 million simulated pure-birth phylogenies with taxon sampling (black) and 9005 post burn-in trees (red) for Cytb  and Rag1 . Hatched red line indicates values for MCC tree.Click here for fileA list of currently recognized North American OPM cyprinid species, including GenBank accession numbers for previously published and newly sequenced data utilized in this study. Benthic/pelagic designations are based on a number of morphological and ecological characters, and references for these characters are given below the table.Click here for file


Respiratory motion compensation is essential for reproducible and robust cardiovascular MRI. Traditionally, breathholds or prospective gating by bellows or navigator signals limit data acquisition to the quiescent phase of respiration . These aFive healthy volunteers were imaged on a 3.0T system  using PC VIPR prescribed over a large chest imaging volume . Interleaving of radial projections that traverse the center of k-space allow for retrospective sorting in a flexible fashion. Our traditional retrospective ECG gating was expanded to incorporate additional sorting of the data into respiratory phases based on the bellows signal Fig.  to proviRepresentative flow waveforms for the DAo during inspiration and expiration are shown in Figure A 3D radial double-gated acquisition and reconstruction that allows for blood flow analysis over the cardiac cycle is presented. Data acquired in healthy volunteers shows significant increased blood flow during expiration compared to inspiration in the Ao. The same effect is marginal in the DAo. Acquisition throughout the respiratory cycle does not prolong scan time while allowing for retrospective selection of arbitrary measurement planes. Further studies are warranted to quantify these respiratory effects in other vessels and to investigate implications for assessing flow.NMSS fund RC1003-A-1, NIH grant 2R01HL072260, and GE Healthcare


Achieving tolerance or drug minimization after transplantation and thus preventing permanent immunosuppression with all the known severe side effects is the most important goal in transplantation medicine. In the last 20 years major progress has been made in understanding the tolerance underlying mechanisms and develop therapeutic strategies in small animal models. However, such knowledge could be rarely translated into the development of successful new therapeutic approaches in clinical transplantation. The success is limited by clinical challenges which are not present in our clean animal facilities such as 1) heterologous immunity - pathogen-specific memory T and B cells recognize alloantigens and boost the immune response towards the allograft, 2) pre-sensitization of recipients - presence of allo-specific memory T and B cells which are inert to most known therapeutic regimens. Thus we know now that we need more personalized treatment strategies according to the patient's immune reactivity. Such a strategy should combine three important aspects: i) an improved immune monitoring; ii) treatment which target memory cells and iii) strategies to reinforce regulation.We have established preclinical transplant models with preformed allo-reactive or pathogen-specific memory T cells in which we compare effectiveness of different treatment approaches combining depletional with regulatory approaches.Furthermore, we have performed a DNA microarray screen on samples of transplant patients and identified surface molecules specifically expressed by naïve, central memory, effector memory or terminal differentiated effector memory (TEMRA) T cells. Using this approach we hope to develop antibodies, which specifically deplete effector memory and TEMRA cells but spare naïve and central memory T cells. Such a treatment will be associated with less side effects e.g. infectious complications as compared to global depletion of T and B cells.


An original total ankle replacement design was developed with the aim of establishing compatibility between the prosthetic articulating surfaces and the retained ligaments. This was achieved with a special shape of a conforming meniscal bearing, free to move forwards/backwards on both metal components during dorsi/plantar flexion. Careful kinematics analyses were carried out in patients after this replacement to assess the functional performance during activity of daily living. A thorough assessment shall include standard gait analysis (GA) and the more accurate motion tracking of the components by 3D fluoroscopic analysis (FA).Eleven patients implanted with the BOX Ankle  were analyzed at 12 months after surgery. GA was performed during stair-climbing/descending using a 8-cameras motion system , electromyography , and an established protocol for lower limb joint kinematics and kinetics [Nearly physiological joint kinematic patterns were observed in both legs Table . A statiNearly normal kinematics and kinetics at the main joints were observed also at the replaced leg. In addition, nearly natural function was restored at the replaced ankle, with large coupled motion in the three anatomical planes. The meniscal-motion was coupled with ankle flexion, supporting the main claims of the designers.


CaseusJM1, a 936-type phage isolated from an Irish dairy plant.The 936-type lytic bacteriophages are the most frequently encountered species infecting lactococcal dairy starters. Infection by members belonging to this species has a significant negative impact on the cheese production process. Here we report the complete genome sequence of the bacteriophage  Lactococcus lactis represent important starter cultures in the dairy industry, and despite decades of research, such starter strains continue to suffer from phage attacks  FLX titanium sequencer . A 110-fold sequencing coverage was obtained using pyrosequencing technology on a 454 FLX instrument. The files generated by the 454 FLX instrument were assembled with GSassembler  to generate a consensus sequence. Several PCR products covering various sections of the genome were sequenced to ensure correct assembly and the resolution of any remaining base conflicts occurring within homopolymer tracts.The sequencing of the http://www.sanger.ac.uk/resources/software/artemis/). Each of the determined ORFs was functionally annotated using BLASTp (Open reading frames (ORFs) were automatically predicted using Heuristic Approach for Gene Prediction . Proteing BLASTp .CaseusJM1 was found to have a double-stranded linear genome of 30,692 bp with a G+C content of 34.84%. A total of 51 ORFs were predicted and were divided into a modular organization based on previously published 936-type phages (CaseusJM1 exhibits a high level of homology to genomes of other 936-type phages (>85% identity). Continued analysis of the genomic organization and diversity is essential to help understand why these phages represent such a significant and constant threat to the dairy industry.e phages  with earCaseusJM1 has been deposited in GenBank under accession number KC522412.The complete genome sequence of lactococcal phage


Panicum virgatum, and mixed-grass prairie) not currently managed as crops to act as post-breeding and fall migratory stopover habitat for birds. In total, we detected 41 bird species, including grassland specialists and species of state and national conservation concern . Avian species richness was generally comparable in switchgrass and prairie and increased with patch size in both patch types. Grassland specialists were less abundant and less likely to occur in patches within highly forested landscapes and were more common and likely to occur in larger patches, indicating that this group is also area-sensitive outside of the breeding season. Variation in the biomass and richness of arthropod food within patches was generally unrelated to richness and abundance metrics. Total bird abundance and that of grassland specialists was higher in patches with greater vegetation structural heterogeneity. Collectively, we find that perennial biomass feedstocks have potential to provide post-breeding and migratory stopover habitat for birds, but that the placement and management of crops will be critical factors in determining their suitability for species of conservation concern. Industrialization of cellulosic bioenergy production that results in reduced crop structural heterogeneity is likely to dramatically reduce the suitability of perennial biomass crops for birds.Increased production of biomass crops in North America will require new agricultural land, intensify the cultivation of land already under production and introduce new types of biomass crops. Assessing the potential biodiversity impacts of novel agricultural systems is fundamental to the maintenance of biodiversity in agricultural landscapes, yet the consequences of expanded biomass production remain unclear. We evaluate the ability of two candidate second generation biomass feedstocks (switchgrass,  Panicum virgatum) or mixed-grass prairie In North America, land-use changes associated with the expansion of contemporary bioenergy crops are generally expected to reduce biodiversity in affected regions Perennial feedstocks can attract a number grassland bird species during the breeding season Our goal is to directly address this information gap by comparing the bird communities in two important candidate biomass feedstocks with potential to provide post-breeding and migratory stopover habitat: Switchgrass and mixed-grass prairie. We first ask if feedstocks differ in the species richness, species density (species richness per unit area) and abundance of migratory bird communities they support, and then investigate how food availability and habitat structure and composition at multiple spatial scales  shape the distributions of birds during the fall migratory period. Because the post-breeding and en-route habitat requirements of grassland birds are poorly-known, we base our predictions about grassland bird responses to crops on established bird-habitat relationships during the breeding and wintering periods. Grassland bird diversity during the breeding season has been linked to plant species diversity P = 0.69), species density  or abundance  was evident across study plots.We identified 95.1% of the 979 individuals detected within transects to the species level. In total, we detected 41 bird species, with greater total and obligate species richness in mixed-grass prairie  than switchgrass  and had a lower forb composition than prairie reconstructions . These aWe detected several species of state and continental conservation concern in both feedstock types . Raw speThe high energetic demands of migration predict that food availability should be an important component of habitat quality for migrants and those individuals preparing to migrate The richness of migratory bird communities increases with patch size in forested systems Many grassland specialist birds are ‘area-sensitive’  during the breeding season Grassland specialist birds commonly avoid selecting breeding habitat within highly forested landscapes Instead, top models of species density and abundance were positively linked with decreasing cover of low, open habitat types and increasing urban land cover. Other studies linking migrant diversity and abundance to surrounding land cover composition are lacking, but this result is surprising given that species richness of breeding bird species decline in relation to increasing urbanization at local and landscape scales Assessing the biodiversity impact of novel production systems is fundamental to reconciling the demands of biodiversity conservation and agricultural production Results of this study suggest that the latter effect is likely to dramatically reduce the suitability of biomass crops as autumn stopover habitat. Switchgrass and prairie are generally expected to be harvested in September, but are somewhat flexible. Harvest strategies that create within-crop structural diversity (e.g. strip harvesting) or that produce a mosaic of harvested and unharvested patches during the migratory period could be a useful management tool Fifteen sites of each of the two biomass crop treatments were selected from established patches throughout southern Michigan . We visiMelospiza melodia) We surveyed the bird community associated with crop patches in the fall of 2008 and 2009, making three visits to each patch: 1) Sep. 7–Sep. 15, 2) Sep. 23–Oct 1, and 3) Oct 7–16. While this time period corresponds to the period of peak migratory movement for migrant land birds in this region Ammodramus spp.). Observations of individuals not identified to the species level were used only to estimate community-wide abundance. Bird surveys were conducted during the first four hours after sunrise.Fixed-width transects In order to obtain representative samples of bird communities in patches differing in area without pseudo-replicating To estimate patch-scale species richness, we used area searches to survey portions of each patch not covered by transects. To maintain observer effort proportional to the size of each patch, observers walked at a regular pace though each patch in a systematic pattern such that one observer passed within 75 m of every point in a patch exactly once. Species detected during strip-transect surveys, including those detected at a distance of >50 m, were pooled with detections from area searches to provide an estimate of bird species richness within each patch.During the second site visit we characterized vegetation structure of crops within each 100 m-long transect to determine how microhabitat gradients may affect spatial distributions of birds. We randomly selected five non-overlapping sampling points within each transect at which we recorded vertical density of vegetation and canopy coverage. Vertical density (an index of biomass) Settlement behavior in grassland birds is frequently linked to landscape composition at larger spatial scales (1000–1600m) We used principal components analysis to reduce the number of within-patch vegetation structural and landscape-scale variables at the 0.5 km and 1.5 km scales into component variables. We employed an orthogonal rotation method that minimizes the number of variables with high loadings on each axis. Microhabitat variables were moderately correlated  and we eArthropod food availability has been linked to the distributions of post-breeding and migratory birds We tested for spatial autocorrelation among sites by comparing residuals of bird community models by using the Moran's index (I) as a function of spatial distance We took a model selection approach to determine the relative importance and effect size of 11 environmental variables and feedstock type  in explaWe modeled the richness of the avian community using generalized linear models with a Poisson distribution and log-link function using SPSS version 15 a priori candidate models that reflected our assessment of likely causes of variation in richness, species density, occurrence and abundance. Our analyses included models of each explanatory variable alone, and two- and three-variable models that we determined to be ecologically relevant. Because species-area relationships may differ by feedstock, we also included models with interactions between patch size and feedstock type. We evaluated the degree of support for logistic models using Akaike's second-order information criterion with a small sample size adjustment (AICc) c (quasi-AICc) which accounts for potential overdispersion of generalized linear models c or ▵QAICc values and normalized Akaike weights (iw). We considered models with ▵AICc or QAICcc or QAICc>2 and We developed a set of Table S1Fit of global models for avian community metrics in model selection analyses. The fit of generalized linear models is assessed as c-hat. A c-hat approximating 1 indicates good fit, while a value greater than 1, but less than 4 indicates moderate to severe overdispersion (DOCX)Click here for additional data file.Table S2Correlation matrix of microhabitat vegetation structural and composition variables. Microhabitat variables describing the structure and composition of biofuel crop stands were moderately-correlated.(DOCX)Click here for additional data file.Table S3Eigenvalues of the first three orthogonal microhabitat principal components extracted.(DOCX)Click here for additional data file.Table S4Loading matrix for the first microhabitat principal component. The principal component was positively related to vertical density and percent grass cover, and negatively related to forb cover.(DOCX)Click here for additional data file.Table S5Correlation matrix of land-use categories in landscapes surrounding focal patches at the 0.5 km scale. The percent cover of forest in landscapes surrounding biofuel crops was negatively correlated with crop cover, urbanization and open habitat types at the 0.5 km scale.(DOCX)Click here for additional data file.Table S6Eigenvalues of the first four orthogonal landscape principal components extracted at the 0.5 km scale.(DOCX)Click here for additional data file.Table S7Loading matrix for the first two landscape principal components at the 0.5 km scale. Landscape principal component 1 exhibited a strong positive relationship with the cover of cropland and open habitats, while landscape principal component 2 was most strongly characterized by a strong positive relationship with urbanization.(DOCX)Click here for additional data file.Table S8Correlation matrix of land-use categories in landscapes surrounding focal patches at the 1.5 km scale. The percent cover of forest in landscapes surrounding biofuel crops was negatively correlated with crop cover while urban cover and open habitat types were also negatively correlated at the 1.5 km scale.(DOCX)Click here for additional data file.Table S9Eigenvalues of the first four orthogonal landscape principal components extracted at the 1.5 km scale.(DOCX)Click here for additional data file.Table S10Loading matrix for the first two landscape principal components at the 1.5 km scale. Landscape principal component 1 exhibited a strong positive relationship with forest cover and a strong negative relationship with crop cover, while landscape principal component 2 exhibited a strong negative relationship with urbanization and a positive relationship with open habitats including old fields, prairie, switchgrass and pasture.(DOCX)Click here for additional data file.


Congenital bicuspid aortic valve (BAV) is the commonest inherited cardiac defect that is often associated with a medley of other cardiovascular anomaly, the most frequent being aortic dilatation which may be a consequent of an interplay between genetic and hemodynamic factors. This retrospective study aims to find a correlation between BAV valve morphology with thoracic aortic dimension with cardiovascular magenetic resonance imaging (CMR).A retrospective analysis of the aortic valve and aortic (Ao) dimension was made of 149 patients aged between 6 and 77 years  with BAV who underwent CMR study. BAV patients with associated coarctation or valvular dysfunction were excluded. Images were acquired with either a Siemens Avanto or Sonata 1.5T scanner in orthogonal planes. Aortic maximal intraluminal dimensions in end diastole were measured at the annulus, sinuses, sino-tubular junction, mid ascending aorta, arch and proximal descending aorta. These measurements were indexed according to body surface area (BSA). BAV valve morphology were characterised in cross-sectional steady-state free precession (SSFP) cine images and grouped according to the presence or absence of a raphe.Of the valve morphology, majority of patients (59.7%) had a raphe, almost always seen between the fused right and left cusps (89%). The indexed aortic root and ascending aortic dimensions were found to be smaller in patients with remnant raphe BAV as compared with those without a raphe (‘pure' BAV). There was a trend towards smaller indexed proximal aortic arch dimensions in BAV patients with valvular raphe although this did not reach statistical significance (p=0.07). There were no difference in indexed aortic arch and proximal descending aortic dimension between the 2 groups as shown in the table below:Majority of BAV patients have a remnant raphe, commonly between the fused right and left cusps. These patients were also found to have smaller indexed aortic root and ascending aortic dimensions compared with those with ‘pure' BAV which may indicate an inherent difference in biologic aortic structure.No funding


In order to differentiate Epstein-Barr virus (EBV) virion DNA versus viral DNA released from tumor cells, we have taken advantage of the observation that viral episomal genomes of herpesviruses are methylated in latently infected cells whereas un-methylated genomes are synthesized and packaged into virions during the lytic phase. We used paramagnetic beads linked to methylCpG binding protein to separate virion and cell-derived viral DNA.DNA isolated from EBV Figure  virions Tumor derived viral DNA can be distinguished from virion associated viral DNA based on preferential binding to methylCpG binding protein. Tumor derived viral DNA was predominantly present in the blood from patients with Hodgkin-Lymphoma, but not in patients without EBV associated malignancy. This technique may be applied to detect tumor derived viral DNA in the blood of patients with EBV associated malignancies.


The literature data demonstrate that the initial stage of abdominal sepsis (AS) is characterized with typical immune shifts as white blood cell (WBC) activation, decrease of T-lymphocyte and B-lymphocyte function followed by dysimmunoglobulinemia. As AS progresses immunity deficiency acquires severe combined character. Impact of immunity deficiency on a course is not described.Escherichia coli K30 LPS was used as antigen for AE antibody detection. All data were compared with healthy donors (99 patients). As a method of immunity correction we selected Sandoglobulin , introduced as 3 ml once intravenously.We investigated 33 patients  with abdominal sepsis . Anti-endotoxin (AE) antibodies  were determined by original modification of hard-phase immunoenzyme analysis. n = 6, no evidence from normal parameters) and low initial immunity . All patients needed comprehensive medication support. The patients with high immunity did not need any immunotherapy. But the patients with low initial immunity required immune therapy to avoid severe course of peritonitis and unfavorable outcomes. Single introduction of Sandoglobulin on the fifth day after surgery was accompanied with increase of immunity indices . Successful surgical treatment and immunotherapy of abdominal sepsis are accompanied by activation of AE immunity, all-class anti-LPS-immunoglobulin concentration growth that blocks mechanisms of further inflammation progress and result in rapid patients' recovery.According to results of immunity parameter investigation on the day of admission we shared all patients into two groups: with high initial immunity (es Table . SimultaPeritonitis patients with initial low immunity need passive nonspecific immunotherapy that stimulates organism protective functions and promotes rapid recovery.


Ferrets are widely used as animal models for studying influenza A viral pathogenesis and transmissibility. Human-adapted influenza A viruses primarily target the upper respiratory tract in humans (infection of the lower respiratory tract is observed less frequently), while in ferrets, upon intranasal inoculation both upper and lower respiratory tract are targeted. Viral tropism is governed by distribution of complex sialylated glycan receptors in various cells/tissues of the host that are specifically recognized by influenza A virus hemagglutinin (HA), a glycoprotein on viral surface. It is generally known that upper respiratory tract of humans and ferrets predominantly express α2→6 sialylated glycan receptors. However much less is known about the fine structure of these glycan receptors and their distribution in different regions of the ferret respiratory tract. In this study, we characterize distribution of glycan receptors going beyond terminal sialic acid linkage in the cranial and caudal regions of the ferret trachea (upper respiratory tract) and lung hilar region (lower respiratory tract) by multiplexing use of various plant lectins and human-adapted HAs to stain these tissue sections. Our findings show that the sialylated glycan receptors recognized by human-adapted HAs are predominantly distributed in submucosal gland of lung hilar region as a part of O-linked glycans. Our study has implications in understanding influenza A viral pathogenesis in ferrets and also in employing ferrets as animal models for developing therapeutic strategies against influenza. The host tissue or cell tropism of the viruses have been investigated previously using Ferrets are widely used as an animal model for understanding influenza A viral pathogenesis and transmission Sambucus nigra agglutinin (SNA-I) and Maackia amurensis lectin (MAL-II) One of the important factors governing the tissue or cellular tropism of the virus is the specific binding of its surface glycoprotein hemagglutinin (or HA) to sialylated glycan receptors  on the host cell surface The advancements in the chemical and chemoenzymatic synthesis of glycans have led to development of microarray platforms comprising of hundreds of diverse glycan structures and structural motifs displayed on various surfaces Given the importance of ferrets as widely used animal models for influenza infection, in this study, we have systematically characterized the glycan structural motifs distributed in the tracheal and hilar regions (primary sites of influenza A virus infection) of the ferret respiratory tract - on any N-linked or O-linked glycan, thereby displaying the broadest specificity to α2→6 sialylated glycans. Therefore staining pattern of SNA broadly indicates distribution of α2→6 sialylated glycans. In order to understand more specifically the distribution of receptors for human-adapted influenza A viruses, recombinantly expressed HAs of two prototypic human-adapted pandemic virus strains from the 1918 H1N1 pandemic, A/South Carolina/1/1918 (or SC18), and from 1957–58 H2N2 pandemic, A/Albany/6/58 (or Alb58) were used. While SC18 has been shown to specifically bind to at least tetrasaccharide glycan motifs terminated by α2→6-linked sialic acid, Alb58 shows a broader specificity by recognizing a minimum trisaccharide motif . MAL-II was used to characterize distribution of avian receptors. As observed earlier, human-adapted viruses showed predominant binding to non-ciliated (including goblet cells) cells in the upper respiratory tract The lectins and recombinant HAs were used individually or in a multiplexed fashion (co-staining) to delineate glycan structures in regions of the ferret respiratory tract including cranial and caudal parts of the tracheal tissue section and lung hilar section. These regions are known to be infected by human-adapted influenza A viruses. Based on the staining patterns, a detailed picture of the distribution of sialylated glycan structural motifs was obtained that goes beyond terminal sialic acid linkage. The extent of lectin staining of various cell types in the different tissue sections was visually scored and is summarized in Paraffinized tissue sections (5 µm thickness) were stained with hematoxylin and eosin to identify distinct cell types and compare it with that of human respiratory tract. Tracheal tissue sections from the cranial and caudal regions and lung hilar region were analyzed . HoweverMaackia amurensis agglutinin (MAL-II) was used to probe α2→3 sialylated glycan receptor distribution in ferret respiratory tract  and from the 1958 pandemic (Alb58) show efficient aerosol transmission in ferrets SC18 HA showed predominant staining of submucosal glands in both ferret tracheal and lung hilar section  and lung hilar sections . All the plant lectins and HAs used in this study showed a substantial loss of staining upon Sialidase A treatment, thereby confirming that staining of tissues by lectins is due to their specific binding to sialylated glycans on the tissue sections .In order to verify the sialic acid binding specificity of lectins used in this study, the tissue sections were treated with Ferrets have been used extensively as a model to study influenza A virus transmission and pathogenesis. Upon intranasal inoculation of the virus, ferrets exhibit similar clinical manifestation as that of humans although there seems to be a difference in viral tropism between ferrets and humans. Upper respiratory tract is the primary site of influenza A infection in humans. Apart from upper respiratory tract, involvement of lower respiratory tract (lung hilar region) is also reported in ferrets. Viral tropism is determined by distribution of influenza A glycan receptors, which are recognized by the viral hemagglutinin (HA) during infection.in vitro binding assay to determine the pattern of viral attachment (PVA) of fluorescein labeled human and avian influenza A viral strains to tissues  has been performed Plant lectins such as SNA-I have been used to stain ferret tracheal tissues in the past and it is generally known that these tissues predominantly express α2→6 sialylated glycan receptors similar to human tracheal tissues Recently, with the emergence of glycan array technology, the glycan binding specificities of several plant lectins and influenza A virus hemagglutinins have been extensively characterized. In this study we used a combination of both lectins and recombinant human-adapted HA to systematically stain both the upper and lower respiratory tissues  in ferrets. The use of recombinant human-adapted HAs and co-staining with multiple lectins permitted us to define glycan receptors for human-adapted influenza A viruses going beyond terminal sialic acid linkage and map their distribution parts of the ferret respiratory tract Figure 6Sialidase A treatment, our results suggest that goblet cell region in human tracheal tissue might have a more predominant expression of the sialyl-Tn-antigen motif than the submucosal region in the ferret lung hilum although both these regions appear to be target sites for binding by human-adapted influenza viruses. This result is also supported by recent findings that show increased expression of sialyl-Tn antigen after virus infection due to goblet cell and/or acinar gland neoplasia in ferrets Our observations support the notion that the receptors for human-adapted influenza A viruses in ferrets are O-linked α2→6 sialylated glycans  that are predominantly distributed in the submucosal glands of the lower respiratory tract (lung hilar region). This notion is also consistent with earlier reports of viral antigens being predominantly found in the submucosal glands in ferret trachea and lung hilar region upon infection with human influenza A virus It is interesting to note that though glycan receptors for human influenza A viruses are differentially distributed in humans and ferrets, they are predominantly expressed in the context of O-linked glycans in either goblet cells (in humans) or in submucosal glands (in ferrets). Given that these O-linked glycans are predominantly expressed in the context of mucins, our study provides a basis to further investigate the role of mucins in influenza A infection. We speculate that by infecting the mucin secreting cells such as goblet cells and submucosal glands, human-adapted influenza A viruses can be easily encapsulated into respiratory droplets formed during sneezing that can in turn facilitate efficient airborne transmission of the virus. In fact, the efficient transmission via respiratory droplets is a hallmark property of human-adapted viruses in the ferret animal models In summary, using a panel of lectins, we have systematically characterized the glycan receptor distribution for influenza A HA in ferret upper and lower respiratory tract, especially receptors relevant to human adapted viruses Figure 6WT HA gene, was used to infect (MOI = 1) suspension cultures of Sf9 cells  cultured in BD Baculogold Max-XP SFM . The infection was monitored and the conditioned media was harvested 3–4 days post-infection. The soluble HA from the harvested conditioned media was purified using Nickel affinity chromatography . Eluting fractions containing HA were pooled, concentrated and buffer exchanged into 1× PBS pH 8.0 (Gibco) using 100K MWCO spin columns . The purified protein was quantified using BCA method (Pierce).Briefly, recombinant baculoviruses with Sialidase A pretreatment, tissue sections were incubated with 0.2 units of Sialidase A  for 3 hours at 37°C prior to incubation with the proteins. Sections were then washed and viewed under a Zeiss LSM510 laser scanning confocal microscope.Formalin fixed and paraffin embedded ferret tracheal and lung hilar tissue sections were obtained from Lovelace Respiratory Research Institute and Division of Comparative Medicine (Massachusetts Institute of Technology) respectively. The lung hilar tissue sections were obtained from 4 year old male normal ferrets that were not exposed to prior influenza A virus infection. The tracheal sections were also obtained from normal ferrets that were not exposed to prior influenza A virus infection. Tissue sections were deparaffinized, rehydrated and pre-blocked with 1% BSA in PBS for 30 minutes at room temperature (RT). HA-antibody pre-complexes were generated by incubating 20 µg/ml of recombinant HA protein with primary  and secondary  antibodies in a ratio of 4∶2∶1 respectively for 20 minutes on ice. Tissue binding studies were performed by incubating tissue sections with the diluted HA-antibody complexes for 3 hours at RT. To visualize the cell nuclei, sections were counterstained with propidium iodide  for 20 minutes at RT. In the case of Formalin fixed and paraffin embedded ferret tracheal and lung hilar tissue sections were obtained from Lovelace Respiratory Research Institute and Division of Comparative Medicine (Massachusetts Institute of Technology) respectively. Tissue sections were deparaffinized and rehydrated. The tissue sections were incubated with 10 µg/ml of lectins  respectively for 3 hours in dark at RT. Lectins were obtained from Vector Labs. To visualize the cell nuclei, sections were counterstained with propidium iodide  for 20 minutes at RT. Sections were then washed, mounted and viewed under a Zeiss LSM510 laser scanning confocal microscope. For MAL-II, since the lectin was biotinylated, prior to adding the lectin, the tissue sections were incubated with streptavidin/biotin kit (Vector Labs) to block endogenous biotin for preventing non-specific staining of the tissues. For lectin co-staining experiments, Jacalin and SNA-I (10 µg/ml each) was added simultaneously to the tissue sections.Table S1Glycan binding specificities of lectins used in this study. Shown in the table is the panel of lectins used in this study and the cartoon representation of glycan motifs recognized by these lectins. The “{“ used to indicate that the glycan motif on the left of “{“ can be linked to either one or more branching positions on the N-linked core glycan structure or different O-linked core glycan structures. Glycan cartoon representation key: N-acetyl-D-neuraminic acid (purple diamond), D-galactose (yellow circle), D-mannose (green circle), N-acetyl-D-glucosamine (blue rectangle), N-acetyl-D-galactosamine (yellow rectangle).(PDF)Click here for additional data file.Figure S1Submucosal gland co-staining with Jacalin and Alb58 HA. Submucosal glands in the ferret trachea showed extensive co-staining with Jacalin and Alb58 HA. The co-staining is indicated by a yellow staining pattern. The submucosal glands are marked by white dotted circle. The apical surface is marked with a white arrow.(PDF)Click here for additional data file.Figure S2Sialic acid-binding specificity of lectins.Sialidase A from Arthrobacter ureafaciens was used to cleave all the sialic acids from the ferret lung hilar tissue sections prior to staining with SNA-I, Alb58 HA and SC18 HA (green). A substantial reduction in staining was observed upon Sialidase A treatment which indicated sialic acid specific binding of HA and lectins in these tissue sections. The apical surface is marked with a white arrow.(PDF)Click here for additional data file.


Mutagenic DNA repair (MDR) transiently increases mutation rate through the activation of low-fidelity repair polymerases in response to specific, DNA-damaging environmental stress conditions such as ultraviolet radiation (UVR) exposure. These repair polymerases also confer UVR tolerance, intimately linking mutability and survival in bacteria that colone habitats subject to regular UVR exposure.Pseudomonas cichorii 302959. Relative fitness measurements of isolates and population samples from replicate lineages indicated that adaptive improvements emerged early in all lineages of our evolution experiment and specific increases in relative fitness correlated with distinct improvements in doubling and lag times. Adaptive improvements gained under UVR and non-UVR conditions were acquired preferentially, and differentially contributed to relative fitness under varied growth conditions.Here, we investigate adaptive specificity in experimental lineages of the highly UVR-mutable epiphytic plant pathogen P. cichorii 302959.These results support our earlier observations that MDR activation may contribute to gains in relative fitness without impeding normal patterns of adaptive specificity in  Mutation is the most fundamental source of variation on which natural selection may drive evolution. However, the predominantly deleterious nature of mutation maintains selection pressure in favor of lower mutation rates as evidenced by the conservation of multiple DNA error-avoidance and error-repair processes Escherichia coli DinB) and polV (E. coli UmuDC) rescue stalled replication forks and permit DNA synthesis across damaged regions of DNA in a template-independent manner by nature of their high processivity and low fidelity Inducible mutability in the form of mutagenic DNA repair (MDR) transiently increases mutation rate through the activation of low-fidelity repair polymerases in response to specific environmental stress conditions such as ultraviolet radiation (UVR) exposure. UVR directly damages DNA causing lesions, including cyclobutane pyrimidine dimers and 6-4 photoproducts, that distort the helical structure, interrupting replication fork progression and inducing the SOS response. The SOS regulon coordinates the control of more than 40 unlinked genes involved in DNA repair, recombination, and cell cycle control Pseudomonas cichorii 302959 rulAB, a homolog of umuDC found in various plant-associated Pseudomonas species P. cichorii 302959 were maintained in a serial transfer regime for 500 generations during which half received daily MDR activation in the form of UVR exposure MDR-mediated UVR tolerance provides a critical ecological advantage to epiphytic plant pathogens harboring Y family polymerases that reside in habitats optimized for solar UVR exposure The results of our initial study suggested that regular activation of inducible mutability in the form of MDR was not detrimental to fitness, but rather could contribute to adaptation and genetic diversity P. cichorii 302959 by examining physiological components of relative fitness improvements. Relative fitness measurements indicate an overall growth advantage that we hypothesized could be subdivided into discrete improvements in bacterial physiology to reveal a more detailed understanding of the adaptation of P. cichorii 302959 to specific laboratory environments. Our results suggest that improvements in doubling and lag times contributed to specific relative fitness gains by both UVR and non-UVR lineages in their respective environments. Surprisingly, specific fitness improvements appeared relatively early in all experimental lineages, independent of regular MDR activation.In the present study, we investigated adaptive specificity in experimental lineages of P. cichorii 302959 was originally isolated in Japan P. cichorii 302959 were derived from a single ancestral colony and propagated by serial transfer for 500 generations in minimal liquid media −2 of UVC (254 nm) radiation from an XX-15 UV lamp  that resulted in ∼10% survival by each lineage population. The energy output of the lamp was monitored with a UV-X radiometer fitted with a UV-25 sensor  and determined to be 1.5 J m−2 s−1. Following UVR irradiation, cultures were diluted 1∶100 into fresh medium and incubated under dark conditions to minimize photoreactivation. These lineages diversified into a fluctuating coexistence of ‘round’ (R) and ‘fuzzy’ (F) colony morphotypes. Eight non-UVR lineages (numbered 33–40) were diluted daily into fresh medium without MDR activation by UVR exposure and contained only the R morphotype. The culture transfer strategies for both UVR and non-UVR lineages resulted in a 1000-fold daily increase in the growth of each population, representing ∼10 generations of binary fission.The ancestral ‘round’ strain of e.g., 25R refers to an isolate from lineage 25 that exhibits the round colony morphotype). Each lineage contained ∼1010 total P. cichorii 302959 cells at stationary phase, and the relative fitness of these communities has been shown to rely on the ecological interactions of sympatric genotypes 8 cells thawed from preserved UVR lineages to maintain the assemblage of diverse genotypes. Isolates and population samples from generation 500 characterized here are identical to those reported previously Samples from each lineage were periodically preserved in a nonevolving state in 10% glycerol at −80°C. All evolved isolates in this study were derived from the ancestral genotype following either 250 or 500 generations of selection in the lineage experiment . We defiP. cichorii were cultured at 28°C in Luria–Burtani (LB) broth , in Davis Minimal broth supplemented with 25 mg l−1 glucose (DM25) (Difco), or on King's medium B (KB) agar −1, gentamicin 10 µg ml−1. Competent cell preparation was performed according to Choi et al.et al.Strains of xylE) which causes expressing colonies on agar media to turn yellow when sprayed with 0.1 M catechol. The relative fitness (W) of the two competitors was calculated as the ratio of their realized growth rate as described previously W = 1.The relative fitness of all isolates and population samples was determined by direct competition with the ancestor under both UVR and non-UVR conditions to identify patterns of specific or general adaptation as described previously t-tests against the hypothesis of mean equal fitness (W = 1). Fitness measurements were compared using two-tailed paired t-tests to determine any significant differences in adaptation between growth intervals (generations 250 vs. 500) or growth environments (UVR vs. non-UVR conditions).The six replicates of each relative fitness measurement were analyzed by two-tailed, independent Before optical density (OD) measurement, lineage isolates and population samples were grown overnight in LB broth and subcultured into 10 ml DM25 at 28°C for 24 hr acclimation. Cultures were diluted 1000-fold into fresh DM25, and 100 µl samples were removed for OD measurement at 600-nm in 96-well microtiter plates using a Tecan Safire . Samples were removed for measurement at 0 hr and at 1 hr intervals from 4 hr to 15 hr. Growth curves for each lineage isolate and population sample were repeated in triplicate under both UVR and non-UVR conditions.600 values from growth curves were standardized by dividing by the initial OD600 (0 hr) and then log2 transformed. The transformed values were plotted, and the window of exponential growth was identified by linear regression ODN1 and N2 are raw, untransformed OD values corresponding to the window of exponential growth observed in the log2 transformation plot and t is the time interval between them where −2 UVC as described above. Cell suspensions were mixed continuously while receiving UVR doses to eliminate survival as a result of shading. Following irradiation, surviving cells were enumerated by dilution plating conducted under dark conditions.Lineage isolates and population samples were grown overnight in LB broth and 2 ml of culture were pelleted, washed with 1 ml saline, resuspended in 1 ml saline, and held on ice. The cell suspensions were mixed with 9 ml saline in a glass petri dish and exposed to a single dose of approximately 140 J mP. cichorii 302959 ancestor under both UVR and non-UVR conditions to examine adaptation during experimental evolution. We evaluated the relative fitness of each lineage by competing both population samples (a subset of the entire lineage maintaining the assemblage of diverse genotypes) and single-colony isolates. In our previous study, we analyzed changes in relative fitness in replicate lineages after 500 generations The relative fitness of all lineages was determined by competition with the P>0.320). Therefore, measurements of the two groups have been combined when comparing isolates from UVR and non-UVR lineages but are also presented individually to provide additional sampling of the genetic diversity present in UVR lineages. Relative fitness measurements were also conducted using population samples to observe any influence of genotype community structure in UVR lineages. We have combined the relative fitness values determined at generation 250 in this study with previously published measurements at generation 500 t-test against the hypothesis of mean fitness equal to that of the ancestral P. cichorii 302959 (W = 1) and is significant where indicated in the figures. Paired t-tests were also used to determine significant differences in relative fitness between growth environments (UVR vs. non-UVR conditions) or growth intervals (generations 250 vs. 500) and these P values are listed at the bottom of each figure.Isolates from UVR lineages expressed two different morphologies, R and F, and single-colony representatives of each from generations 250 and 500 were characterized. Neither morphotype group exhibited a significant fitness advantage under either UVR or non-UVR conditions  with the exception of the isolate from lineage 37 . When compared to isolates from generation 500, there was no significant difference in relative fitness under non-UVR conditions  were also observed. The ancestral tion 500 . Lag timtion 500 . The lagtion 500 . Converstion 500 . A compl−2 s−1 was used to resolve subtle differences in UVR tolerance that could not be distinguished at the lower dosage used during lineage propagation. The ancestral P. cichorii 302959 strain exhibited a 0.7% survival rate at the elevated dose. Population samples from UVR lineages displayed the most consistent improvements in UVR tolerance with average survival rates of 4.3% and 4.0% at generations 250 and 500, respectively. R and F morphotype isolates exhibited a mixture of UVR tolerance phenotypes that were higher, lower, or comparable to that of the ancestor. A complete list of UVR tolerance measures for each population sample and isolate is available in Changes in UVR tolerance could lead to changes in relative fitness under UVR conditions. Therefore, the UVR tolerance of population samples and isolates from each UVR and non-UVR lineages taken at generations 250 and 500 was determined following a single dose of UVC. An elevated dose of ∼140 J mP. cichorii 302959 ancestor under both non-UVR and UVR conditions. The resulting fitness trajectories are plotted in To track changes in relative fitness during lineage growth over the course of 500 generations, preserved population samples from four randomly selected lineages were obtained at different intervals and competed against the Similarly, non-UVR lineages 36 and 39 exhibited fitness gains under their native, non-UVR conditions first . NeitherP. cichorii 302959 emerged early and correlated with discrete growth improvements but was not dependent upon MDR activation. In a previous study, we maintained parallel cultures of P. cichorii 302959 for 500 generations with daily MDR activation in the form of UVR exposure and observed higher relative fitness particularly under UVR conditions P. cichorii 302959 to evaluate this observed adaptive specificity.Our results suggest that adaptive specificity in laboratory evolved populations of the highly UVR-mutable P. cichorii 302959 lineages emerged within the first 250 generations of growth. Additional fitness gains attained between generations 250 and 500 were primarily achieved under UVR conditions, even in non-UVR lineages. The dramatic changes in fitness exhibited by F isolates from lineages 27 and 28 demonstrate that UVR lineage populations contained a dynamic balance of genotypes, and that isolates sampled at generation 500 were not necessarily derived from those sampled at generation 250. Together, these results emphasize the complexity of adaptation in the UVR lineage populations due to the high adaptive potential and plasticity of the ancestral P. cichorii 302959 inducible mutator genotype. Additionally, the increase in relative fitness observed only under UVR conditions between generations 250 and 500 in some non-UVR lineages offers the first indication that beneficial mutations differentially contribute to fitness under the two growth conditions.The relative fitness of all 16 lineages was determined at the generation 250 midpoint of our evolution experiment with the intent to gain further information on the acquisition of fitness observed previously at the generation 500 endpoint. Fitness measurements at the midpoint can offer some indication of the timing, number, magnitude, and parallel nature of adaptive improvements achieved by each lineage. Measurements of relative fitness at generation 250 reflected the same patterns of adaptive specificity observed previously at generation 500 P. cichorii 302959 obtained at generations 250 and 500 under both UVR and non-UVR conditions. Growth dynamic improvements correlated well with changes in relative fitness such that nearly all evolved lineages with increased fitness displayed corresponding reductions in both doubling and lag times compared to the ancestor. The average reductions in both lag and doubling times exhibited by UVR lineages were greatest under UVR conditions while non-UVR lineages displayed comparable reductions under both conditions. These results are consistent with the observed patterns of adaptive specificity in relative fitness measurements suggesting that specific growth improvements are responsible for the characteristic gains in relative fitness displayed by UVR lineages under UVR conditions.Measures of relative fitness indicate the overall advantage of one bacterial strain or population over another but provide no information about the underlying physiological differences responsible for the advantage. Changes in growth dynamics have been identified as distinct components of relative fitness improvements in similar studies conducted previously Improvements in lag time could be indicative of a faster metabolic transition from stationary phase to exponential growth due to adaptation to the culture media or to the cyclic nature of the serial transfer regime. Improvements in lag time under UVR conditions specifically could result from increased UVR tolerance or more efficient DNA repair in the form of improved SOS induction or polymerase processivity. The complex SOS regulatory network is comprised of both transcriptional and post-transcriptional regulators that tightly control the temporal activity of its gene products E. coli, adaptive mutations that contribute to survival through catabolism of cellular detritus P. cichorii 302959 or any evolved lineages in glucose-free Davis Minimal medium amended with a bacterial suspension killed either by prolonged heat or UVR exposure (data not shown). Therefore, increased relative fitness under UVR conditions and the adaptive specificity of UVR lineages that is reflected in improved growth likely did not result from either increased UVR tolerance or a heightened ability to scavenge nutrients as a carbon source.We explored changes in UVR tolerance as an obvious candidate for improved fitness and lag time under UVR conditions but only observed increased UVR tolerance concurrently with fitness gains in a limited number of instances. In general, there was no pattern to suggest that greater UVR tolerance was primarily responsible for increased relative fitness observed under UVR conditions. Alternatively, improved fitness under UVR conditions could result from enhanced nutrient scavenging. Approximately 90% of the population died as a result of UVR exposure during each cycle of the UVR lineage regime. The death of these individuals could release additional nutrients in the form of cell lysate that is transferred with surviving cells to fresh medium for the next cycle of growth. Any mutation that would enhance the utilization of these nutrients could lead to greater fitness exclusively under UVR conditions. In evolution experiments with We previously observed the reproducible emergence of an F morphotype that coexisted with the ancestral, R morphotype in all UVR lineages but never in non-UVR lineages P. cichorii 302959 genotype. Furthermore, the early fitness improvements by UVR lineages are consistent with the emergence of F morphotypes and spontaneous rifampicin-resistant colonies reported previously It is clear from our relative fitness measurements that adaptive changes in both non-UVR and UVR lineages were concentrated in the first 250 generations, confirming both the strong selection pressure of the experimental environments and the high adaptive potential of the ancestral P. cichorii 302959 exhibited by both UVR and non-UVR lineages. Such adaptive specialization can result in fitness trade-offs due to either antagonistic pleiotropy or mutation accumulation resulting in lower fitness under different environmental conditions The fitness trajectories in this study also suggest that adaptive improvements differentially contributed to relative fitness gains under UVR and non-UVR conditions. The ordered acquisition of improvements in the two environments favored the respective conditions under which each lineage was propagated such that UVR lineages first exhibited specific fitness gains under UVR conditions and non-UVR lineages under non-UVR conditions. Based on our previous observations of relative fitness at generation 500, we concluded that non-UVR lineages exhibited comparable fitness gains under UVR and non-UVR conditions due to their adaptation to the shared culture medium P. cichorii 302959 genotype is more poorly adapted to the UVR conditions and therefore more adaptive opportunities exist than under non-UVR conditions. Although the first two possibilities lend support to the proposed positive influence of inducible mutability on adaptive evolution, we do not have sufficient information to distinguish them from each other or from the third possibility.Interestingly, when fitness did increase in the alternate environment, corresponding improvements in fitness were not observed in the native environment. It is unlikely that fitness improvements under UVR and non-UVR conditions are independent. Rather, mutations that contribute to fitness observable only in the alternate environment may represent general improvements of smaller effect in the native environment. After gaining fitness in the alternate environment, UVR lineages continued to gain fitness specifically under their native conditions while non-UVR lineages did not, producing the illusion of adaptive specificity exclusively in UVR lineages. We can propose three possible explanations for this discrepancy: (1) MDR activation provides access to more beneficial mutations by altering the mutation spectrum of UVR lineages; (2) by increasing the overall mutation rate, MDR activation increases the rate at which beneficial mutations appear in the population and non-UVR lineages would gain additional relative fitness if propagated for a longer time; or (3) the ancestral P. cichorii 302959 during experimental evolution by measuring relative fitness and corresponding changes in physiology. Our results suggest that adaptive specificity in this organism correlates with discrete growth improvements but does not depend on MDR activation. Adaptive improvements in all lineages were concentrated in the first 250 generations of experimental evolution and specific increases in relative fitness correlated with distinct improvements in doubling and lag times. Furthermore, UVR lineages exhibited additional gains in fitness after generation 250 exclusively under UVR conditions that were reflected in further doubling and lag time improvements but likely not greater UVR tolerance or scavenging of nutrients to support the growth of new biomass. Fitness trajectories of select lineages clearly indicate that adaptive improvements under UVR and non-UVR conditions were acquired preferentially and differentially contributed to relative fitness under varied growth conditions.In summary, we have investigated the adaptive specificity of The results of this study lend support to our earlier observations that suggest increased mutation rate in the form of inducible mutability does not impede adaptation by mutation accumulation. Rather, UVR and non-UVR lineages preferentially acquired adaptive growth improvements in a similar manner and additional fitness gains by UVR lineages may have been due to greater access to beneficial mutations. Our future work will compare the influence of different mechanisms of mutability on adaptation in an advanced genetic system equipped with genomic tools for analysis.Table S1P. cichorii 302959 during growth under non-UVR and UVR conditions.Doubling time of population samples and isolates from lineages of (DOCX)Click here for additional data file.Table S2P. cichorii 302959 during growth under non-UVR and UVR conditions.Lag time of population samples and isolates from lineages of (DOCX)Click here for additional data file.Table S3P. cichorii 302959 following ∼140 J m−2 UVC.Percent survival of population samples and isolates from lineages of (DOCX)Click here for additional data file.


Phocarctos hookeri), a nationally critical, declining species impacted by trawl fisheries. We used satellite telemetry and linear mixed effects models to investigate sex differences in the foraging ranges of juvenile NZ sea lions. Male trip distances and durations were almost twice as long as female trips, with males foraging over the Auckland Island shelf and in further locations than females. Sex was the most important variable in trip distance, maximum distance travelled from study site, foraging cycle duration and percent time at sea whereas mass and age had small effects on these characteristics. Our findings support the predictions of the niche divergence hypothesis, which suggests that sexual segregation acts to decrease intraspecific resource competition. As a consequence of sexual segregation in foraging ranges, female foraging grounds had proportionally double the overlap with fisheries operations than males. This distribution exposes female juvenile NZ sea lions to a greater risk of resource competition and bycatch from fisheries than males, which can result in higher female mortality. Such sex-biased mortality could impact population dynamics, because female population decline can lead to decreased population fecundity. Thus, effective conservation and management strategies must take into account sex differences in foraging behaviour, as well as differential threat-risk to external impacts such as fisheries bycatch.Sexual segregation  is observed in a large range of taxa. Investigating causes for sexual segregation is vital for understanding population dynamics and has important conservation implications, as sex differences in foraging ecology may affect vulnerability to area-specific human activities. Although behavioural ecologists have proposed numerous hypotheses for this phenomenon, the underlying causes of sexual segregation are poorly understood. We examined the size-dimorphism and niche divergence hypotheses as potential explanations for sexual segregation in the New Zealand (NZ) sea lion ( Diomedea exulans) have larger overlap with fisheries activities than males, resulting in higher bycatch mortality of females Sexual segregation, defined here as differential space and resource use by males and females Cervus elaphusLoxodonta africanaPicoides pubescensParus atricapillusNumerous hypotheses have been advanced to explain sexual segregation Anguilla rostrataArctocephalus forsteriThe causes of sexual segregation have been predominantly studied in terrestrial vertebrates Phocarctos hookeri) provides an ideal study system to examine two of the main competing hypotheses for sexual segregation: the size-dimorphism (sex-related size differences) or niche divergence (decreasing intraspecific resource competition) hypotheses. Adult male and female NZ sea lions are thought to forage in different areas Notodarus sloanii) around the subantarctic Auckland Islands, both through mortality in bycatch and potential resource competition The New Zealand (NZ) sea lion (Our study aimed to answer three main questions: (1) What are the foraging ranges and at sea movements of juvenile NZ sea lions at the Auckland Islands? (2) Are the size-dimorphism and niche divergence hypotheses potential explanations for sexual segregation in the foraging ranges of NZ sea lions? Under the size-dimorphism hypothesis, although males are larger and would require more energy per unit time to meet energy requirements, females require higher quality (i.e. higher energy) prey due to the allometric relationship between metabolic requirements and body size/gut capacity This study was conducted with approval from the Department of Conservation (DOC) and University of Otago Animal Ethics Committees . Instrument deployments were performed under full inhalant gas anaesthesia, and all efforts were made to minimize pain and suffering.We collected data over four austral summers from January-February, 2007–2010 at Sandy Bay, Enderby Island in the NZ subantarctic Auckland Islands  and also vehicle-borne high frequency  transmitters to facilitate recaptures. We attached the instruments to the dorsal pelage of the animal below the shoulder blades on the back midline using two-part epoxy resin. Once the instruments were securely attached, we stopped the flow of anaesthetic and the sea lion was allowed to recover. We observed each animal until it was fully conscious and had returned to the location of capture. Where possible, we recaptured the animals before the end of the field season (February 18) to retrieve instruments.Animal positions at sea were calculated by the Argos satellite system and detailed information on the satellite location classes and number of locations received is provided in . We filtAfter filtering satellite data with the state-space model, we defined complete foraging trips as trips with the start and end locations at Sandy Bay or within 10 km of Enderby Island following Chilvers 2008a. We restricted calculations of mean distance travelled per trip to complete foraging trips. Maximum distance travelled from Enderby Island per foraging trip was measured in ArcGIS 9.3.1 The squid fisheries operate between January-May on the Auckland Island shelf, utilising mid-water column and bottom trawls. To assess the potential sex differences in the overlap of juvenile NZ sea lion foraging grounds with the squid trawl fisheries, we mapped the 50% and 95% kernel UDs of combined fishing effort from 2007–2010 (NZ Ministry of Fisheries), along with the kernel UDs of the male and female satellite locations from 2007–2010. To quantify individual sea lion overlap with fisheries, we mapped the 50% and 95% kernel UDs of combined fishing effort from 2007–2010 (NZ Ministry of Fisheries) along with UDs of individual female and male juvenile NZ sea lions.C) scores. Where necessary, the response variables were power or square root transformed to improve the normality of the model residuals. To improve the interpretability of regression coefficients, we centred and standardized the age and mass predictor variables Although inter-annual variation may influence the foraging behaviour observed, small sample sizes per year precluded the differentiation of annual and individual differences. Furthermore, foraging studies on individual adult female NZ sea lions followed across several years indicate no annual variation in foraging behaviour C) scores To identify the relative importance of the predictor variables  across the different foraging trip characteristics and to generate weighted coefficient estimates, we used a model averaging approach based on the Akaike Information Criterion correction for small sample size .From January–February, 2007–2010, we deployed a total of 33 satellite tags on 19 female (2–3 years-old) and 14 male (2–5 years-old) juvenile NZ sea lions . Two malP<0.001). Male trip distances and maximum distance travelled from the study site were almost twice as long as female distances (2 vs. 523 km2), while the 95% kernel UDs of males were over two times larger than females , maximum distance from study site  and foraging cycle duration . Significant repeatability estimates indicate male juvenile NZ sea lions consistently travelled further distances for longer periods than females.Sex had the most important effect on trip distance, maximum distance from study site, foraging cycle duration and percent time at sea  and S3. 2 vs. 3844 km2). As a percentage of the 95% kernel UD area, female foraging grounds had proportionally almost double the amount of overlap with squid trawl fisheries operations than males  with squid trawl fisheries. From 2007–2010, the squid fisheries operated at depths 98–338 m (NZ Ministry of Fisheries). Sixty-one percent of tagged female and 47% of male juvenile NZ sea lions dives were within the depth range targeted by squid trawl fisheries . Based on the assumption that male and female NZ sea lions behave similarly in fishing areas (e.g. one sex does not avoid trawl nets more than the other sex), a consequence of the sexual segregation in the foraging ranges of juvenile NZ sea lions is the higher risk of resource competition and bycatch of females in the squid fishery. Juvenile female NZ sea lion foraging grounds had proportionally double the overlap with fisheries operations than males . Of the Sex-biased mortality of females in fishery operations have been reported for several species  from 2007–2010. The Auckland Islands are represented in light grey. Bathymetric contours are shown as black lines. The Auckland Island shelf is represented by the 500 m bathymetric boundary.(TIF)Click here for additional data file.Figure S2Utilisation distributions (UD) of individual female juvenile New Zealand sea lions (Phocarctos hookeri) and squid trawl fisheries. Individuals with: A) 0–25%, B) 26–50% and C) 51–75% of their 95% UD overlapping with fisheries. Note panel C is represented by two figures of five individual females per map.(TIF)Click here for additional data file.Figure S3Utilisation distributions (UD) of individual male juvenile New Zealand sea lions (Phocarctos hookeri) and squid trawl fisheries. Individuals with: A) 0–25%, B) 26–50% and C) 51–75% of their 95% UD overlapping with fisheries.(TIF)Click here for additional data file.Table S1Satellite location data for female and male juvenile New Zealand sea lions (Phocarctos hookeri).(DOC)Click here for additional data file.Table S2Results of linear mixed effects models run on juvenile New Zealand sea lion (Phocarctos hookeri) foraging trip characteristics: trip distance and maximum distance from study site.(DOC)Click here for additional data file.Table S3Results of linear mixed effects models run on juvenile New Zealand sea lion (Phocarctos hookeri) foraging trip characteristics: foraging cycle duration and percent time spent at sea.(DOC)Click here for additional data file.


To the Editor: Birge and Sonnesyn report the first death of a Minnesota resident caused by Powassan virus (POWV) (1]) .The 17 POWV infections detected in Minnesota residents from 2008 through 2011   were found through enhanced surveillance. Health alerts to Minnesota medical providers described POWV as a possible etiologic agent for viral meningitis and encephalitis. Providers consulted with MDH on suspected cases and submitted serum and cerebrospinal fluid specimens to MDH. MDH conducted serologic testing for endemic arboviruses (including POWV) and reverse transcription PCR (RT-PCR) for flaviviruses and POWV. MDH would not have detected any POWV infections without enhanced surveillance. Limited field studies also identified POWV-infected ticks in 4 Minnesota counties  is preferred (Ixodes scapularis ticks (blacklegged ticks), the tick species associated with deer tick virus transmission, and viruses from all POWV-positive tick pools were confirmed as deer tick virus by sequencing. The distribution of the 2 lineages in North America is poorly understood, and most cases likely go undetected without specific POWV surveillance efforts.Few POWV infections are identified by lineage (prototype vs. deer tick virus); Minnesota’s first case in 2008 was identified as a deer tick virus infection, but the lineage was unknown for the other 16 cases. However, many case-patients had likely exposure to


Fibrotic burden of LV myocardium quantified by CMR provides additional information about LA size and mechanics than morphological severity of LV hypertrophy measured by LV mass index alone.Progressive myocardial stiffness of the LV in patients with hypertrophic cardiomyopathy (HCM) impedes early diastolic filling and left atrial (LA) emptying functions. Over time, LA of patients with HCM progressively dilates and undergoes structural and functional remodeling with serious cardiac events such as development of atrial fibrillation and diastolic heart failure despite optimal medical therapy.We studied 65 HCM patients  with CMR. The fibrotic content of the LV myocardium  was obtained using serial R1 mapping by cine-IR method before and up to 30 minutes after contrast injection. LA volumes and LA emptying function across 3 phases of the cardiac cycles, and LV function were also quantified. We sought to determine the strongest variables that predispose the patients to have LA dilatation and impaired LA emptying function.FIMeanSeg demonstrated strong positive correlation with LA volumes across all 3 phases of the cardiac cycle, with the strongest correlation with LAMax volume at end-ventricular systole (r=0.41, P = 0.0005, Figure FIMeanSeg provides independent association with LV volume beyond knowledge of ventricular functional parameters and severity of ventricular hypertrophy by LV mass. This novel method may add to the current characterization of impaired diastolic function in HCM patients and may have prognostic implications.Yucheng Chen got the salary from China Scholarship Council.


Patients with transposition of the great arteries (TGA) after atrial baffle switch operation show differences in ventricular torsion and outflow tract geometry compared to healthy volunteers. We sought to investigate if these differences in cardio-mechanics translate into abnormal blood flow patterns in the pulmonary trunk and the aorta.Blood flow patterns were assessed with flow-sensitive four-dimensional velocity-encoded magnetic resonance imaging (4D VEC MRI), using a 1,5T Phillips MRI system. Measurements were made in the pulmonary trunk and the aorta in healthy volunteers (n=7) and TGA patients after atrial baffle switch operation (n=10). Blood flow was analyzed for vortex formation using custom-made software.There were clear differences in blood flow patterns between healthy volunteers and TGA patients in both the pulmonary trunk and the aorta. In healthy volunteers, flow was laminar, parabolic in the pulmonary trunk and showed left helical flow pattern in the aorta. In TGA patients we observed opposite flow patterns with predominant parabolic flow in the aorta but helical flow and vortex formation in the pulmonary trunk.There are abnormal flow profiles in the aorta and pulmonary trunk in TGA patients. The data of this study provides evidence that differences in left and right ventricular cardio-mechanics directly translate into different flow patterns.