Candida boidinii, which involve lipid droplets and peroxisomes, are summarized. The physiological functions of Atg (Autophagy-related) proteins in these organisms are discussed in relation to the dynamics of these two important organelles.Eukaryotic microorganisms resident in the phyllosphere  undergo dynamic changes in nutrient conditions and adapt their metabolic pathways during proliferation or in the course of infection of host plants. Some of these metabolic switches are accomplished by regulation of organelle abundance. Recent studies have shown that autophagy plays a major role in reducing the organelle quantity, thereby contributing to the metabolic switch required for survival or virulence of the microorganisms in the phyllosphere. In this mini review the metabolic pathways in several phytopathogenic fungi and the non-infectious asporogenous yeast  Figure 1A). The process of differentiation requires induction/inactivation of specific metabolic pathways. In the following section of this mini review, we first introduce the melanin biosynthesis pathway, which is conserved among a subset of phytopathogenic fungi and is closely associated with the dynamics of lipid droplets and peroxisomes. Then several functional links between this biosynthetic pathway and autophagy are discussed.The phyllosphere has been under evaluated as an ecological site for plant-microbe interaction, compared with the rhizosphere . HoweverCandida boidinii, which can grow on methanol as a sole carbon and energy source , which is vital for the decrease of peroxisome quantity and for the growth of this organism in the phyllosphere, is summarized.Recent studies revealed that non-phytopathogenic eukaryotic microorganisms can also establish an inhabitation in the phyllosphere. One such example is the asporogenous methylotrophic yeast y source . AlthougColletotrichum and Magnaporthe species, develop appressoria pigmented with melanin for efficient infection of their host plants . Consistent with this, it was shown that functional assembly of peroxisomes was required for the formation of melanized appressoria as well as the pathogenicity of Colletotrichum orbiculare . Molecular details underlying this feedback regulation are not clear, but it is possible that the intermediates of the pathway, namely free fatty acids, acetyl-CoA, and/or malonyl-CoA, might act as signaling molecules to repress lipolysis, in order to avoid the deleterious effect called “lipotoxicity,” which results from excess accumulation of lipid substances, especially free fatty acids. In many experimental systems including yeast, the formation of lipid droplets is strongly suggested to contribute to the prevention of lipotoxicity by incorporating free fatty acids into the core contents of the organelle, or into neutral lipids (Our recent study demonstrated that the liberation of fatty acids from lipid droplets (termed lipolysis) in the appressorium was inhibited by blocking either the (1) beta-oxidation pathway, (2) conversion of acetyl-CoA to malonyl-CoA, or (3) consumption of malonyl-CoA by melanin biosynthesis . This inl lipids .Magnaporthe oryzae and Colletotrichum orbiculare; M. oryzae demonstrated that lipid droplets are incorporated into the vacuolar portion of the cell and are degraded therein  and dihydroxyacetone synthase (Das), when these organisms are grown on methanol . Owing tde novo membrane biogenesis . Growth was dependent on its methanol-metabolizing enzymes, Aod1 and Das1. In addition, methanol concentrations available to the organism on the plant leaves were also determined with the fluorescence intensity of Venus expressed under the regulation of a methanol-inducible promoter. Interestingly, dynamic oscillation of methanol concentration was observed on the surface of A. thaliana leaves during the daily light–dark cycle, with higher concentrations in the dark period . In accordance with this oscillation, peroxisome abundance in the inoculated Candida boidinii cells was found to fluctuate as revealed by fluorescence microscopy of a peroxisome-targeted Venus: the organelle quantity increased in the dark period, and decreased in the light period . We assume that the methanol-induced peroxisome serves as a storage organelle for proteins to replenish amino-acid pools needed in the natural environment, in order for the immotile microorganism to survive until they obtain nutrients for further proliferation.Our recent study took advantage of the expression of a fluorescent protein (Venus) for the enhanced, quantitative detection of a leaves . ThroughCandida boidinii possessed a homolog of P. pastoris Atg30 (the pexophagy-specific protein), we inoculated the Cbatg30Δ strain on plant leaves. The decrease in peroxisome quantity in the light period was suppressed by the loss of Atg30. Notably, this mutant strain exhibited a severe growth defect on plant leaves, similar to mutant strains incapable of methanol metabolism. The transport of Atg8 into the vacuole, a marker of general autophagy was detected in the inoculated Candida boidinii Cbatg30Δ strain, but not in the Cbatg1Δ strain, indicating that general autophagy occurs throughout the day. However, it is still unknown how autophagic activity and selectivity are regulated along the daily light–dark cycle.Since Despite great advances in the understanding of the molecular mechanism of pexophagy, the physiological role of this selective autophagy had been an enigma for a long time, until the microorganisms were transferred from laboratory media to plant-surfaces or plant surface-mimicking environments. Likewise, the dynamics of lipid droplets is now an exciting topic of cell biology, and studies of eukaryotic microorganisms in the phyllosphere will give us profound insight into the physiological importance of organelle dynamics. Accumulating molecular information on the microorganisms in the phyllosphere will also be important for a better understanding of plant-microorganism interactions.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Arabidopsis thaliana the gynoecium is composed of two carpels that are fused into a tube-like structure. As the gynoecial primordium arises from the floral meristem, a specialized meristematic structure, the carpel margin meristem (CMM), develops from portions of the medial gynoecial domain. The CMM is critical for reproductive competence because it gives rise to the ovules, the precursors of the seeds. Here we report a functional role for the transcription factor PERIANTHIA (PAN) in the development of the gynoecial medial domain and the formation of ovule primordia. This function of PAN is revealed in pan aintegumenta (ant) as well as seuss (seu) pan double mutants that form reduced numbers of ovules. Previously, PAN was identified as a regulator of perianth organ number and as a direct activator of AGAMOUS (AG) expression in floral whorl four. However, the seu pan double mutants display enhanced ectopic AG expression in developing sepals and the partial transformation of sepals to petals indicating a novel role for PAN in the repression of AG in floral whorl one. These results indicate that PAN functions as an activator or repressor of AG expression in a whorl-specific fashion. The seu pan double mutants also display enhanced floral indeterminacy, resulting in the formation of “fifth whorl” structures and disruption of WUSCHEL (WUS) expression patterns revealing a novel role for SEU in floral meristem termination.The gynoecium is the female reproductive structure of angiosperm flowers. In  Arabidopsis thaliana, as with most angiosperms, reproductive competence depends on the proper development of the flower. Arabidopsis flowers develop from floral meristems, specialized structures that contain organized groups of undifferentiated cells that give rise to the four types of floral organs: sepals, petals, stamens, and carpels  encodes a MADS-domain containing protein that plays at least two key roles during floral genesis: participation in regulatory complexes that specify the identity of stamens and carpels; and bringing about termination of the floral meristem and thus generating a determinant floral structure  in the cells that will give rise to the stamens and carpels    , encodes a transcriptional adaptor protein, that physically interacts with several MADS domain proteins including APETALA1 (AP1), SEPALLATA3 (SEP3), AGAMOUS-LIKE24 (AGL24) and SHORT VEGETATIVE PHASE (SVP) , another transcription factor, to regulate the expression of downstream genes critical for the formation of ovules  allele was previously characterized as a RNA null allele via in situ hybridization to inflorescence and floral tissues  through the 3′ end of cDNA. The antisense WUS probe is complementary to the entire WUS cDNA clone and is derived from BamHI cut pMHwus16 plasmid, a gift of Jenn Fletcher. The AG sense strand control probe was generated from a linearized pCIT565 plasmid (cut with XhoI) using the Sp6 promoter and contains the full length cDNA sequences. To generate the PAN antisense probe, the plasmid G50929 (ABRC) a full-length sequence-confirmed ORF/cDNA clone  for clearing and mounting on slides for visualization. Slides were examined with an Axioscop2 microscope (Zeiss) with Nomarski optics. Ovule counts were made from stages 11–14 gynoecia fixed on slides. Analysis of carpel bending and splitting was done under dissecting scope. Gynoecia were rated from 1 to 4 independently for bending as well as splitting. A severity score was given based on the following scoring system: 1, no defect; 2, mild defect; 3, moderate defect; 4, severe defect. All gynoecia were scored by the same individual, at the same time without knowledge of the genotype. All photos were captured with Q Capture software on a 5.0 RTV digital camera . Data analysis was conducted in JMP Pro 10  using multiple pair-wise comparison of the means with a Tukey-Kramer HSD test at an alpha of 0.05 or with a Student's pan mutant plants we characterized three available pan alleles  ovules per gynoecium  that we tested under our long-day growth conditions . We then used this severity index to generate a mean severity score for the comparison of genotypes of interest. Carpel bending and carpel splitting phenotypes were not observed in the Col-0 gynoecia that we assayed. Under the long-day growth conditions, the g Figure . However Figures , 3A. Thig Figure , the cars Figure .seu pan 057190 showed statistically significant enhancement in carpel bending compared to the pan single mutant (seu single mutants rarely display a bending phenotype under long-day)  Figure . The benseu pan gynoecial defects were enhanced relative to the long-day conditions  To analyze the seu pan phenotypes under short-day conditions we scored the gynoecia for the occurrence of four phenotypes; complete loss of ovules, carpel bending, severe loss of valves , and indeterminate growth from internal gynoecial positions. We did not observe any of these phenotypes in the Col-0 plants and they were found infrequently (5%) in seu or pan single mutants  of the seu pan double mutants displayed a severe alteration in the morphology of the external valve cells  and reduced production of pollen from anthers (data not shown). Additionally, the seu pan double mutant plants displayed an enhanced delay in the transition from vegetative to reproductive development as determined by counting the number of rosette leaves formed before bolting  by repressing the expression of WUS, a stem cell maintenance gene within the floral meristem . Thus, although we were able to document cases of perdurant WUS expression, our data suggests that this ectopic expression is likely relatively short in duration and that WUS expression is not continuously maintained in the fifth whorl structures.The fifth whorl structures formed in the s Figure  suggestiWUS expression in the center of the shoot apical meristem (SAM) and in early stage floral primordia . In some of the seu pan inflorescence meristems the WUS expressing region was expanded and appeared to be punctuated as if several organizing centers had been formed within the potentially compound inflorescence meristem  of the seu pan whorl one floral organs exhibited partial homeotic transformations based on the cell surface morphologies  that PAN plays a role in the development of the medial domain of the gynoecium and that (2) this role is revealed when the activity of SEU or ANT is compromised. The dependence of the phenotype on the loss of function of SEU or ANT suggests that the function of PAN during gynoecial development is partially overlapping with that ofSEU and ANT. The defects of the seu pan double mutant are similar to, but distinct from the ant pan double mutant, indicating a differential sharing of functions between these three genes.The enhancement of gynoecial defects observed in the pan ant double mutants include an enhanced loss of ovules and enhanced splitting of the gynoecial tube relative to the single mutant parents . This data suggests that SEU and PAN share a function in the regulation of carpel number. In the most severely affected seu pan double mutants, there is a loss of the characteristic morphology of the abaxial valve epidermal cells and these cells develop as cells that are morphologically similar to abaxial replum cells  and pan tousled (tsl) double mutants indicated that PAN also shares overlapping functional roles with TSL and ETT during medial domain development    are also likely to be de-repressed in the seu pan double mutant, although we have not yet confirmed this with in situ hybridization experiments. Ectopic expression of B-class genes in whorl one organs could be caused by ectopic AG expression. Previously lug alleles were shown to condition the partial transformation of whorl one organs to petaloid and stamenoid chimeric organs and ectopic expressions of the B-class genes PISTILLATA (PI) and APETALA3 (AP3) were detected in developing whorl one structures . They propose that the ectopic AG brings about an ectopic expression of the B-Class genes and this is consistent with the identification of PI and AP3 as targets of AG regulation by Gomez-Mena et al. . However, both a et al. .SEU and PAN function in a whorl specific fashion in the regulation of AG transcription or activity. SEU and PAN activities are required for efficient repression of AG in whorl 1 while their activities are required for efficient activation of AG function in whorl four. We propose that the differential action of these proteins is due to whorl specific co-factors or post-transcriptional modifications. The identity of these whorl specific modifiers remains to be elucidated.Our data suggests that Zuniga-Mayo et al. previously suggested a relationship between the proper termination of the floral meristem and the subsequent development of the CMM (Zuniga-Mayo et al., jaiba crabs claw double mutants that display a loss of floral determinacy as well as defects in the development the CMM. Our investigation of the seu pan double mutant further supports this possibility. When ovules develop in the seu pan double mutants, they arise at apical positions within the gynoecium, and thus at a distance from the basally-located fifth whorl structures. Thus, if the floral meristem fails to properly terminate, the cells furthest from the perdurant meristem are more likely to form ovules than those closer to the meristem. This could suggest a gradient of an inhibitor from the floral meristem. However, it is equally likely that a temporal effect explains the difference. As the gynoecium grows from the apex, cells that divided temporally later in development will also be found in more apical positions. Thus, the decay over time of any inhibitory effect of the floral meristem might also result in the formation of ovules only at the apex of the seu pan double mutants.This was based on their analysis of April N. Wynn and Robert G. Franks: conceived and designed the experiments, analyzed the data, and wrote the paper; April N. Wynn, Andrew A. Seaman, Ashley L. Jones and Robert G. Franks: Performed the experiments.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Purpose. Transplantation of stem, progenitor, or precursor cells has resulted in photoreceptor replacement and evidence of functional efficacy in rodent models of retinal degeneration. Ongoing work has been directed toward the replication of these results in a large animal model, namely, the pig. Methods. Retinal progenitor cells were derived from the neural retina of GFP-transgenic pigs and transplanted to the subretinal space of rhodopsin Pro347Leu-transgenic allorecipients, in the early stage of the degeneration and the absence of immune suppression. Results. Results confirm the survival of allogeneic porcine RPCs without immune suppression in the setting of photoreceptor dystrophy. The expression of multiple photoreceptor markers by grafted cells included the rod outer segment-specific marker ROM-1. Further evidence of photoreceptor differentiation included the presence of numerous photoreceptor rosettes within GFP-positive grafts, indicative of the development of cellular polarity and self-assembly into rudiments of outer retinal tissue. Conclusion. Together, these data support the tolerance of RPCs as allografts and demonstrate the high level of rod photoreceptor development that can be obtained from cultured RPCs following transplantation. Strategies for further progress in this area, together with possible functional implications, are discussed. As a group, degenerative diseases of the retina constitute a significant source of visual disability, particularly in the developed world, and yet current therapeutic options are quite limited. For instance, the loss of photoreceptor cells, as seen in the later stages of retinitis pigmentosa and macular degeneration, results in permanent visual deficits for which no restorative treatment is available. Nevertheless, the notion that photoreceptor cells might be replaceable in the therapeutic setting has been given recent support by experimental work in animal models.Work in the rat first showed that transplanted neural progenitor cells could migrate into the host retina, take up residence within the cellular laminae of this tissue, and exhibit morphological signs of integration into the local cytoarchitecture –3. SubseMore recent work has shown that many of the results obtained using progenitor cell transplantation in rodents also apply to other mammalian species, including the Brazilian opossum , the pigPrior work in the pig has shown profuse expression of photoreceptor markers by grafted retinal progenitor cells, yet the full extent of donor cell differentiation was difficult to determine, in part due to limited visualization of donor cell morphology , 10. In Two timed-pregnant NT5 GFP-transgenic sows  were sacThe resulting cultures were defined as RPCs and these cells have been further characterized, as previously reported , 10, 18.Cells were passaged for approximately 4–6 weeks prior to transplantation. The cells were grown as monolayers and dissociated with each passage, but prior to transplantation were allowed to form nascent spherical aggregates, and the smaller aggregates were preferentially collected for use as donor material. The reason for adopting this last approach is that fully dissociated RPCs exhibit poor survival rates following transplantation, whereas large spherical aggregates can be associated with glial differentiation within the sphere core .Recipient animals used in the present experiments were 15 transgenic rhodopsin Pro347Leu transgenic swine (from 2 litters: age 6 and 9 weeks) with a known retinal dystrophic phenotype [6 cells) were injected either as spheres or as single cell suspension through the retinotomy and into the retinal bleb using a 27-gauge silicon-tipped needle. Immediate reflux of some cells into the vitreous cavity was observed in some animals. A small air bubble was placed in the subretinal bleb under the retinotomy to prevent further reflux of cells after withdrawal of the needle. Sclerotomies and conjunctiva were sutured with 7–0 vicryl. Lateral canthal incisions were sutured with 6–0 vicryl. The pigs were examined by ophthalmoscopy 1-2 days after surgery.The transplantation technique was similar to that previously described in pigs –10, 18. The research protocol used was previously reviewed and approved by the Danish Animal Experiment Inspectorate, the North Carolina State University IACUC, and completed in accordance with the ARVO statement for the Use of Animals in Ophthalmic and Vision Research.μm on a cryostat. During the sectioning process, every fifteenth section was examined by epifluorescence microscopy for GFP+ cells, and every tenth slide was stained with Hematoxylin-Eosin (Htx-Eosin).Eyes were enucleated following euthanasia by overdose of intravenous pentobarbital at the termination of the experiment. The survival time for the first litter  was 5 weeks posttransplantation (age 11 weeks), while that of the second litter  was 9 weeks (age 18 weeks). Globes were placed in 4% paraformaldehyde (PFA) for 10–20 minutes. The anterior segment and the lens were then removed and the posterior segment was postfixed for 2 hours in 4% PFA, with subsequent rinsing in rising concentrations of sucrose containing Sörensen's phosphate buffer. A horizontal cut was made which extended from the temporal retinal margin to 2-3 mm nasal to the optic disc, thus comprising the temporal ciliary margin, the area centralis, and the optic disc. The tissues were embedded in a gelatin medium and serially sectioned at 12 The retinal sections were exposed to primary anti-sera see  in a moiSections were frequently stained with chicken anti-GFP , either in order to examine the quality of the GFP expression or to reveal possible downregulated grafted cells. The endogenous GFP expression was always high and enhancing with anti-GFP resulted in blurring of the cell boundaries. The GFP staining did not appear to reveal more surviving cells.Progenitor cells from the fetal porcine retina proliferated rapidly in culture, as previously described , 10, 18.Following transplantation to nonimmunosuppressed juvenile rhodopsin Pro347Leu transgenic pigs, GFP+ donor cells could be readily identified at the site of transplantation . Strong Donor cells were predominantly located in the subretinal space directly adjacent to the RPE layer, while a subset of GFP+ profiles migrated or extended processes into the host ONL. Cells in the subretinal space frequently extended processes perpendicular to the retina or with random orientations, whereas GFP+ processes within the host ONL exhibited predominantly radial orientations . Donor pThere was evidence of widespread differentiation of donor RPCs along the photoreceptor lineage following transplantation to the eye of dystrophic allorecipients. This data included both morphological considerations and marker expression. In H&E stained sections, cells within the graft predominantly exhibited small, densely packed nuclei which closely resembled adjacent photoreceptors of the host ONL . In addiFurther support for photoreceptor differentiation was obtained by analysis of marker expression. Grafted cells showed immunohistochemical evidence for expression of multiple photoreceptor-associated markers. Cells of the grafts, including cells within rosettes, showed expression of recoverin and rhodopsin, with a notable degree of double-labeling for these markers Figures  and 5. TIn terms of marker expression patterns, GFP was evident in many grafted cells but was somewhat less expressed within rosettes and was excluded from the central core of these structures Figures  and 5, aA major challenge faced during the development of therapeutic applications for stem cells is the requirement for reliable tissue-specific differentiation of grafted cells. This is particularly true in the setting of the retina, owing both to the need for molecular transduction of photic stimuli as well as the requirement for a highly organized and optically transparent tissue. Here we show that cultured RPCs are capable of attaining a high level of photoreceptor development following transplantation to the retina of a large mammal. In this model, grafted RPCs differentiated into photoreceptor-like profiles in sufficient quantities and with sufficient structural polarity and cytological affinity to self-organize into rosettes. Marker expression closely followed normal patterns, supporting the conclusion that these donor cells had differentiated into morphological photoreceptors.Previous work with stem , progeniWe found that porcine RPCs expressed photoreceptor markers after transplantation to the subretinal space, yet photoreceptor differentiation was also observed in a grafted cluster of cells that adhered ectopically to the optic nerve head. The unintended location of this cluster is likely a consequence of the misdirection of grafted neurospheres, at or near the time of surgery, presumably due to reflux from the subretinal space through the retinotomy, as is known to occur. In this instance, the potential for confusion with host cells was eliminated by endogenous GFP reporter gene expression. Interestingly, this data supports the concept that RPCs are capable of spontaneously differentiating into photoreceptors at relatively high yield without the need for microenvironmental cues such as might be obtained from engraftment in the host retina. In addition, the photoreceptor cells generated within the graft exhibited sufficient polarity to self-organize into rosettes. This particular cluster contained multiple rosettes of the type typical of those formed by photoreceptor cells after various perturbations of the retina, wherein the nuclei are densely packed around the periphery and the inner and outer segments are oriented inward to form the rosette's core. These types of highly organized rosettes can be viewed as focal tissue rudiments, each attempting to recapitulate the structure of the outer nuclear layer (ONL), albeit in a manner lacking global continuity. Rosettes of this type are to be distinguished from the more primitive Flexner Wintersteiner rosettes seen in a number of neoplastic conditions.The functional capabilities of the grafted cells were not assessed; however, prior work with embryonic retinal transplants in rodents has demonstrated that rosettes of similar morphology are capable of detecting light and providing limited light-mediated behaviors, even in the absence of an RPE layer , 27–29. Appropriately localized expression of the rod-specific marker ROM-1 seen in the current study provides additional evidence of the high level of photoreceptor differentiation obtainable from cultured RPCs. The presence of ROM-1 in the core of rosettes is consistent with the development of rod outer segments (ROS) by the donor-derived photoreceptors. The survival of the grafted cells seen in the current study, in the absence of immune suppression, provides additional confirmation for the high level of tolerance shown to allogeneic RPCs following introduction to both the vitreous cavity and subretinal space of a retinal dystrophic large mammal. Less certain is the contribution of the transgenic rhodopsin Pro347Leu background to the integration of donor cells. RPCs, like many other types of neural progenitor and precursor cells, are known to display an evident tropism for areas of degeneration, trauma, or disease. It was anticipated that the use of dystrophic hosts would result in enhanced intraretinal integration by grafted RPCs. GFP+ profiles did exhibit radially oriented integration into the host ONL and extension of elaborater processes in the orthogonal plane; however, the identity of the integrated cells remains to be determined, as does the question of graft-host connectivity.The pig has emerged a preferred species for modeling of surgical procedures, including ocular applications. The increasing availability of transgenic swine and recent sequencing of the porcine genome provide additional advantages to the use of this model. The current study illustrates the value of the porcine model in the translational development of regenerative strategies and, in particular, intraocular stem cell transplantation.


Balaenoptera musculus spp) are an endangered species found in the Southern and Indian Oceans. Here two recognized subspecies of blue whales and, based on passive acoustic monitoring, four “acoustic populations” occur. Three of these are pygmy blue whale (B.m. brevicauda) populations while the fourth is the Antarctic blue whale (B.m. intermedia). Past whaling catches have dramatically reduced their numbers but recent acoustic recordings show that these oceans are still important habitat for blue whales. Presently little is known about the seasonal movements and degree of overlap of these four populations, particularly in the central Indian Ocean. We examined the geographic and seasonal occurrence of different blue whale acoustic populations using one year of passive acoustic recording from three sites located at different latitudes in the Indian Ocean. The vocalizations of the different blue whale subspecies and acoustic populations were recorded seasonally in different regions. For some call types and locations, there was spatial and temporal overlap, particularly between Antarctic and different pygmy blue whale acoustic populations. Except on the southernmost hydrophone, all three pygmy blue whale acoustic populations were found at different sites or during different seasons, which further suggests that these populations are generally geographically distinct. This unusual blue whale diversity in sub-Antarctic and sub-tropical waters indicates the importance of the area for blue whales in these former whaling grounds.Understanding the seasonal movements and distribution patterns of migratory species over ocean basin scales is vital for appropriate conservation and management measures. However, assessing populations over remote regions is challenging, particularly if they are rare. Blue whales ( Balaenoptera musculus subspp., (Linnaeus 1758) was abundant in the Southern Hemisphere. In only a few decades, the largest living animal was hunted to the brink of biological extinction B.m. intermedia) and the smaller pygmy blue whale (B.m. brevicauda) B.m. indica, by some based on the original description by Blyth (1859). Although there is little morphometric evidence for this subspecies designation, the indica name should have precedence over brevicauda if all these populations are considered to be pygmy blue whales Prior to intensive commercial whaling, the blue whale, It is presumed that during the period of legal whaling in the Southern Hemisphere (1904–1964), most of the blue whales killed were the Antarctic subspecies. Pygmy blue whales were only discovered in 1959–60 Most of our current knowledge of blue whale distribution in the Indian and Southern Oceans is derived from long-term passive acoustic monitoring. Passive acoustic monitoring has been used successfully in many oceans of the world to study the presence, migratory movements and feeding behavior of blue whales e.g. . This meAntarctic blue whales are found largely south of 60°S in the austral summer where they feed, and then likely spread out over a broader range during the austral winter Pygmy blue whales are rare at latitudes south of 60°S Euphausia superba) exploit the presence of abundant phytoplankton near ice edges, continental shelves and dynamic frontal regions Blue whales are known to concentrate in regions of high primary and secondary productivity year-round The Indian Ocean is clearly important habitat for at least four blue whale populations but little is still known about the seasonal movements and degree of overlap of these populations, particularly in the center of the Indian Ocean. In this study, we analyzed more than one year of passive acoustic recording from each of three sites located at different latitudes of the Indian Ocean basin to examine the geographic and seasonal occurrence of different blue whale acoustic populations to better understand their distribution.Three autonomous hydrophone packages were deployed near the French territories in the Southern and Indian Oceans from October 2006 to January and April 2008 Blue whales worldwide produce stereotyped calls with high intensity, low frequency and long duration. They also produce downswept short duration (1 to 4 s) and frequency modulated calls (90 to 25 Hz) named D calls www.birds.cornell.edu/brp/). The Antarctic blue whale call template consisted of one complete call. Only single units of each pygmy blue whale calls were used in those templates because they were usually above ambient noise level. We selected the third unit of the Sri Lanka call; the second unit of the Madagascar call, and the harmonic at 70 Hz of the second unit of the Australian call . In 2007, 4,532 calls were detected at MAD from March through June, with peaks in April and May, while only half as many calls were detected at SWAMS from December to May . Madagascar calls predominantly occurred in autumn . The distribution of call detections by season was quite different between the two sites: at NEAMS nearly 68% of calls were detected in winter while at SWAMS over 70% were detected in autumn  and for relatively short periods of the year. In 2007, four times more calls were detected at SWAMS  than at NEAMS , and over a longer period, January to June versus March to June, respectively . The din autumn . No AustThe vocalizations of the different blue whale subspecies and populations were recorded seasonally in three different areas of the Indian Ocean. For some call types and locations, there were spatial and temporal overlaps, particularly between Antarctic blue whale calls and the different pygmy blue whale calls. Except on the southernmost hydrophone (SWAMS), the different calls of the three pygmy blue whale acoustic populations were found at different sites or during different seasons, further suggesting that these populations of pygmy blue whales preferentially occupy different regions of the Indian Ocean, in addition to being acoustically distinct. Overall, when placed in the context of previous work , the results here show the distribution of subspecies and populations at basin scales and emphasize the importance of the Indian Ocean for blue whales .Historically, Antarctic blue whale distribution has been defined by a migration pattern between high-latitude summer feeding areas and low-latitude wintering areas Sri Lanka blue whales are presumed to be resident in the northern Indian Ocean Madagascar calls were only recorded in autumn and winter months at the westernmost station (MAD) and from late summer through winter months at the southernmost station (SWAMS). This seasonal pattern of Madagascar calls matches all other reports of acoustic detections in the Indian Ocean to date . MadagasAustralia-type calls were recorded at relatively few locations in the Indian Ocean . No AustThere were similarities in the seasonal detections of Australia and Madagascar pygmy blue whale calls, but they appeared to be geographically separated between the eastern and western Indian Ocean basins, respectively, as previously suggested . Both MaBecause we detected individual calls, rather than using an energy ratio method, which detects the acoustic energy from an unknown number of calls, it is likely that the detected calls were from animals that were relatively close to the instruments. In a comparison of the two methods on a same data set, Širovic et al (2009) In the vicinity of the southernmost station (SWAMS), four acoustic populations of blue whales use these sub-Antarctic waters at different times of the year, but sometimes overlap in space and time. Madagascar and Australia pygmy blue whales co-occur at sub-Antarctic latitudes during the end of summer and autumn months. This is similar to the pattern reported in the southwest part of the basin One of the biggest gaps in knowledge in blue whale acoustic ecology is a lack of understanding of the relationship between the number of calls and the number of calling whales. An increase in the number of calls from one month to another may be the result of an increase in the number of calling whales, or that calling whales stay in the detection area during a longer period of time, or that individual whales may increase their call rates seasonally. Regardless, the high number of calls highlights the importance of the sub-Antarctic Indian Ocean for Antarctic blue whales either as a wintering ground or as a final destination before moving southwards to the ice edge of the Antarctic continent.The easternmost station (NEAMS) is roughly 1500 km to the northeast of SWAMS, at the edge of the northern sub-tropical front. Here, the detection patterns of blue whale calls are clearly different. The acoustic diversity was less since no Madagascar pygmy blue whale calls and only few Australia type calls were recorded. However, some Sri Lanka type calls were present here year-round, particularly during austral summer. This area could be a possible wintering ground for this population in addition to that hypothesized for Crozet Islands The northernmost station (MAD) is in sub-tropical waters 2400 km away from the southernmost station and 2500 km west of the easternmost station. The acoustic diversity here is generally poor. This area is not exploited by Australia pygmy blue whales, and perhaps the instrument is at the edge of the distribution of this population. More surprisingly, no Sri Lanka pygmy blue whale calls were detected here; this population may be distributed in a more northerly and easterly manner in the Indian Ocean. Madagascar pygmy blue whales were detected exclusively during autumn months. Unsurprisingly, fewer Antarctic blue whales were detected in these sub-tropical latitudes although the number of calls increased during winter months. During winter, Antarctic blue whales may move northwards and are widely dispersed at different latitudes in the Indian Ocean. However, the year-round presence of calls, especially in this area, far away from the main Antarctic feeding grounds, suggests that some individuals remain at lower latitudes possibly to exploit food resources present in the region. It is also possible that individuals are moving between the Antarctic and sub-Antarctic within a season such that, rather than year-round occupancy of some individuals, there is year-round occupancy of some part of a broader population. The age, reproductive status or individual condition may explain these geographic differences but there are no data to support this at present.Blue whale sightings in the Indian Ocean are rare and difficult to obtain, and long-term passive acoustic recorders provide most of the recent information about the seasonal occurrence and movements of these animals over this remote and large area. As more information from long-term passive acoustic recorders in other regions and over longer time periods is gathered, the importance of the Indian Ocean as year-round habitat for at least four acoustic populations of blue whales, including the Critically Endangered Antarctic blue whale


Tumors can metastasize to the oral cavity and affect the jaws, soft tissue and salivary glands. Oral cavity metastases are considered rare and represent approximately 1% of all oral malignancies. Because of their rarity and atypical clinical and radiographic appearance, metastatic lesions are considered a diagnostic challenge. The purpose of this report is to present a rare case of a metastatic breast carcinoma mimicking a periodontal abscess in the mandible.A 55-year-old Caucasian woman was referred to our clinic for evaluation of bisphosphonate-induced jaw osteonecrosis. She had undergone modified radical mastectomy with axillary lymph node dissection for invasive ductal carcinoma of the left breast. Her clinical examination showed diffuse swelling and a periodontal pocket of 6 mm exhibiting suppuration in the posterior right mandible. Moreover, paresthesia of the lower right lip and chin was noted. There were no significant radiographic findings other than alveolar bone loss due to her periodontal disease. Although the lesion resembled a periodontal abscess, metastatic carcinoma of the breast was suspected on the basis of the patient's medical history. The area was biopsied, and histological analysis confirmed the final diagnosis of metastatic breast carcinoma.The general dentist or dental specialist should maintain a high level of suspicion while evaluating patients with a history of cancer. Paresthesias of the lower lip and the chin should be considered ominous signs of metastatic disease. This case highlights the importance of the value of a detailed medical history and thorough clinical examination for the early detection of metastatic tumors in the oral cavity. Metastases in the oral cavity are rare and comprise approximately 1% of all oral malignancies ,3. The mA wide range of clinical signs and symptoms may be seen in association with metastatic tumors of the oral cavity, with the most common being pain, swelling, altered sensation, halitosis, gum irritation, tooth loosening and mobility, exophytic masses of the soft tissues, trismus and, rarely, pathologic fractures ,2,4. Numet al. [Metastatic tumors of the oral cavity do not exhibit a pathognomonic radiographic appearance; therefore, radiographic examination is rarely considered diagnostically important. Osteolytic radiolucent lesions with ill-defined and irregular margins may be seen, while osteoblastic lesions with a pure radiopaque or a mixed radiopaque-radiolucent appearance are typically associated with prostate cancer ,4,6. EarThe purpose of this report is to describe a rare case of a metastatic breast carcinoma in the mandibular gingival tissue that mimicked a periodontal abscess.A 55-year-old Caucasian woman with subtle pain and tenderness in the area surrounding the right third mandibular molar was referred to our clinic by her oncologist with the provisional diagnosis of bisphosphonate-induced jaw osteonecrosis. Her medical history revealed a modified radical mastectomy with axillary lymph node dissection for invasive ductal carcinoma of the left breast. The tumor was positive for estrogen receptors and cerbB2, but negative for progesterone receptors; thus she received adjuvant hormone therapy with tamoxifen. Moreover, bisphosphonate treatment was initiated with 4 mg of intravenous ibandronic acid administered every three weeks.An intra-oral examination revealed diffuse swelling of the buccal gingiva surrounding the second and third molar teeth that was soft and tender on palpation, with signs of inflammation Figure . The invA panoramic radiograph showed generalized horizontal bone loss throughout the patient's dentition Figure . A peri-On the basis of the patient's medical history and paresthesia of the lower lip and chin, metastatic disease was highly suspected. The differential diagnosis included acute or chronic periodontal abscess, acute alveolar abscess, bisphosphonate-induced jaw osteonecrosis and osteomyelitis.The swelling of the buccal gingiva was biopsied. Five-micron-thick, formalin-fixed, paraffin-embedded tissue sections stained with hematoxylin and eosin showed a fragment of parakeratinized oral mucosa infiltrated by solid and cribriform nests of neoplastic cells in a vascular and myxofibromatous stroma Figure . The neoThe patient was referred back to her oncologist. A full body scan did not reveal additional metastases, and a technetium-99 m-methylene diphosphonate bone scan located a region of increased radioisotope uptake ("hot spot") on the posterior right side of the mandible.The bisphosphonate treatment was continued, and local irradiation of the right posterior mandible was administered as palliative treatment. Although extraction of the involved teeth prior to radiotherapy was feasible, it was decided to preserve them and re-evaluate their prognosis during the follow-up appointments. The patient underwent radiation therapy with a cumulative dose of 3000 cGy fractionated over two weeks, which resulted in complete relief of her symptoms and remission of the disease Figure . Follow-The diagnosis of metastasis to the oral cavity is a significant challenge to the clinician because of the lack of pathognomonic signs and symptoms. To the best of our knowledge, this is the first reported case of a metastatic breast cancer mimicking a periodontal abscess. Previously described cases of metastases to the periodontal tissues were associated with extensive osteolytic destruction of the alveolar bone and root apex resorption -9, even Our patient was referred to our clinic by her oncologist for the evaluation of possible osteonecrosis of the jaw caused by bisphosphonate treatment. The patient's oral cavity was carefully examined, but no signs of exposed avascular necrotic bone were found in the mandible. The existence of exposed necrotic bone over a period of eight weeks with past or recent use of bisphosphonates is an essential element for rendering the diagnosis of osteonecrosis associated with bisphosphonates, along with the absence of previous radiation therapy to the jaws . TherefoThe local inflammation of the soft tissues that surrounded the area and the periodontal pocket exhibiting suppuration were signs of possible inflammatory reactions such as an acute or chronic periodontal abscess, an acute alveolar abscess or a combined endodontic-periodontic lesion. However, the relatively healthy condition of the patient's teeth, the existence of vital pulp after several diagnostic tests and the lack of radiographic signs eliminated the possibility of an endodontic-related lesion.In the case presented herein, the location of the swelling, spontaneous intra-pocket suppuration and the existence of typical signs of periodontal disease were suggestive of a periodontal abscess. Periodontal abscesses are most often associated with a pre-existing periodontal pocket and present as an ovoid elevation of the gingival tissue along the lateral side of the root . DependiParesthesia of the lower lip and chin is the major symptom suggestive of metastatic disease. It is described in the literature as mental nerve neuropathy or numb chin syndrome (NCS) ,15. The The management of metastatic breast carcinomas of the oral cavity is primarily palliative and may include radiotherapy, chemotherapy, hormone therapy and, rarely, surgical intervention. Pain relief and avoidance of possible infections, fractures or hemorrhage should be the major goals . Local rThe prognosis for patients with metastatic lesions of the oral cavity is generally poor, primarily because of the delay in the detection of the lesions. The average survival time for patients with metastatic tumors in the oral cavity is six to seven months, with approximately 70% of patients dying within one year of diagnosis ,19,20. MIn conclusion, this case illustrates the importance of suspecting a metastatic lesion in the jaw, despite the lack of clinical or radiographic evidence. The general dentist or dental specialist should obtain the patient's complete medical history and carefully evaluate unusual clinical and radiographic findings such as lip and chin paresthesias to differentiate metastatic lesions from clinically similar entities. As these lesions are associated with a poor prognosis, early detection is of extreme importance.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.The authors declare that they have no competing interests.PE and MI analyzed and interpreted the patient data. TK performed the histological examination of the biopsy specimen and was involved in the manuscript editing and review. PE was involved in the literature review as well as manuscript preparation, editing and submission. MI was involved in the manuscript editing and review. All authors read and approved the final manuscript.


Escherichia coli chromosome is organized into four macrodomains, the function and organisation of which are poorly understood. In this review we focus on the MatP, SeqA, and SlmA proteins that have recently been identified as the first examples of factors with macrodomain-specific DNA-binding properties. In particular, we review the evidence that these factors contribute towards the control of chromosome replication and segregation by specifically targeting subregions of the genome and contributing towards their unique properties. Genome sequence analysis of multiple related bacteria, including pathogenic species, reveals that macrodomain-specific distribution of SeqA, SlmA, and MatP is conserved, suggesting common principles of chromosome organisation in these organisms. This discovery of proteins with macrodomain-specific binding properties hints that there are other proteins with similar specificity yet to be unveiled. We discuss the roles of the proteins identified to date as well as strategies that may be employed to discover new factors.The  All organisms are faced with the challenge of organising their genetic content within the confines of the cell or its compartments. In eukaryotes, DNA is packed inside the nucleus and histone proteins are known to wrap DNA into nucleosomes. Nucleosomal arrays are folded into chromatin fibers, which are themselves folded into higher order structures. Whilst our understanding of this process at the nucleosomal level is well developed, higher levels of organization are poorly understood E. coli has been established during the last 5 years by Boccard and coworkers att sites scattered throughout the E. coli chromosome att sites that supported efficient recombination and thus were spatially close. On the basis of these observations, it was concluded that the E. coli chromosome is organized into four discrete structured subdomains and that att sites in each domain interact primarily with the att sites in the same domain. Each of these domains  contains approximately 1 Mbp of DNA. The localization of the macrodomains is subject to changes during the cell cycle, but is fairly well defined , which is found 50 times along the E. coli K-12 genome. Strikingly, none of these sites are found in the Ter macrodomain and they are underrepresented in the Left and Right macrodomains , was discovered in silico by searching for scattered domain-specific targets of nucleoid-associated proteins. The factor specifically binding to this site (MatP) was identified in DNA-binding assays using crude E. coli extracts ycbG gene. The high affinity binding of MatP within the Ter domain was visualized in vivo using fluorescent microscopy. These experiments showed that MatP prevents premature chromosome segregation early during the cell cycle by keeping the Ter regions of two chromosomes together. In MatP knock-out cells this prolonged colocalization of the Ter domains is not observed. Fast growing cells deficient in MatP display a filament-like or anucleate phenotype. A delay in segregation of the daughter chromosomes due to the binding of MatP to the Ter region thus appears essential in coordinating chromosome segregation and cell division. Also, without MatP, the Ter domain displays higher mobility and a lower degree of compaction. Surprisingly the effects of MatP-DNA binding stretch over long distances. The deletion of a matS site increases the mobility of regions even several tens of kb away. While the role of this protein in the cell cycle and the organization of the Ter domain is apparent, the mechanism of MatP action is still unknown. Two models have been proposed for how MatP organizes the Ter domain. According to the first model MatP dimers bridge two matS sites located on either separate chromosomes or within one chromosome. It is possible, that bridging nucleates at matS sites and that flanking regions are zipped up by additional nonspecific binding (and bridging) of MatP. The second model invokes an as yet unknown cofactor. After the binding of MatP, this factor would be recruited to regions surrounding matS sites and spread over distances up to several kb. An obvious candidate for such binding would be the H-NS protein MatP is a small DNA-binding protein that—unlike SeqA and SlmA—is associated exclusively with the Ter domain of the i genome  [20]. ItmioC, dnaA, ftsZ, and mukB), SeqA binding is thought to exert cell cycle–dependent control on gene expression seqA mutant As mentioned above, SeqA, SlmA, and MatP are distinct from the classical nucleoid-associated proteins in that they recognise DNA with a high degree of sequence specificity. In this respect the DNA-binding properties of SeqA, SlmA, and MatP are more akin to those of transcription factors. Intriguingly, many SeqA binding sites are located at promoters and within coding regions of genes involved in DNA replication and repair E. coliThe pattern of SeqA, SlmA, and MatP binding is probably similar among Gram negative bacteria, including the many pathogenic organisms, related to


Aspergillus flavus (Link ex Fr.) has been the focus of intensive research due to the production of carcinogenic and highly toxic secondary metabolites collectively known as aflatoxins following pre-harvest colonization of crops. Given this recurrent problem and the occurrence of a severe aflatoxin outbreak in maize (Zea mays L.), particularly in the Southeast U.S. in the 1977 growing season, a significant research effort has been put forth to determine the nature of the interaction occurring between aflatoxin production, A. flavus, environment and its various hosts before harvest. Many studies have investigated this interaction at the genetic, transcript, and protein levels, and in terms of fungal biology at either pre- or post-harvest time points. Later experiments have indicated that the interaction and overall resistance phenotype of the host is a quantitative trait with a relatively low heritability. In addition, a high degree of environmental interaction has been noted, particularly with sources of abiotic stress for either the host or the fungus such as drought or heat stresses. Here, we review the history of research into this complex interaction and propose future directions for elucidating the relationship between resistance and susceptibility to A. flavus colonization, abiotic stress, and its relationship to oxidative stress in which aflatoxin production may function as a form of antioxidant protection to the producing fungus.Since the early 1960s, the fungal pathogen  Aspergillus flavus  by A. flavus is not primarily due to the expression of symptoms known as Aspergillus ear rot but, rather, is due to the subsequent contamination of the grain with the fungal metabolite aflatoxin. Aflatoxins are a group of polyketide-derived furanocoumarin secondary metabolites produced by certain species of fungi, including the genus Aspergillus , lipases, pectinases (P2c), proteases such as alkaline protease, and xylanases , A. flavus.Several constitutively expressed and inducible proteins have been described in the literature, which have been shown to counter the function of other hydrolytic virulence proteins produced by mination . In addi. flavus . In peanA. flavus and aflatoxin contamination on the basis of utilizing phenotypic screenings and molecular markers associated with known avirulence genes to identify quantitative trait loci (QTL) for selection  a large genotype x environment (G × E) interaction conditions the expression of this trait. For example Accumulation of such antifungal and avirulence proteins has been shown to contribute to the resistance observed in several maize lines along with morphological characteristics of resistant kernels such as thickened wax cuticles . Variouselection . These eh others , illustrA. flavus and aflatoxin production, particularly abiotic stresses such as drought and heat stress. This was clearly demonstrated as early as 1977 when widespread and intense drought conditions in the Midwestern and Southeastern United States resulted in a high degree of aflatoxin contamination of maize kernels pre-harvest , and a 17.9-kDa heat shock protein (HSP17.9). A. flavus colonization.It has been hypothesized that there is an underlying relationship among various molecular mechanisms of drought stress adaptation and resistance to A. flavus resistance. The WRKY transcription factors have been shown to regulate the responses of multiple plant species to both biotic and abiotic stresses (ZmWRKY33 was recently shown to enhance abscisic acid (ABA) signaling and enhance osmotic stress tolerance in maize seedlings  is known to induce a response to oxidative stress and regulate the expression of antioxidant enzymes like catalase , herbivory caused significant increases in lipid peroxidation and hydroxyl radical formation in the soybean leaves. The activities of several oxidative enzymes  were also increased following H. zea herbivory on soybean (Glycine max) leaves. They concluded that oxidative responses in the soybean plants may have led to a decrease in herbivory and an increase in oxidative damage to the plant. Diuraphis noxia Mordvilko), and the non-leaf-chlorosis-eliciting bird cherry-oat aphid , which differ in oxidative enzyme activities. While only peroxidase activity was detected in R. padi, catalase activity was only detected in D. noxia. The oxidative responses of four cereal plants  to the feeding of the two species of aphid differed  are critical for the management of pest outbreaks and the reduction of mycotoxin contamination in agricultural crops.A. flavus and aflatoxin contamination. Although this conclusion seems credible due to the high degree of correlative evidence present in the literature, the exact mechanism of how this phenomenon functions has yet to be elucidated completely. More recent studies into the biology of A. flavus and the mechanisms regulating the production of aflatoxin may illuminate this issue , as well as antioxidant enzyme activities, were present in toxigenic strains in comparison to non-toxigenic strains. This indicated that oxidative stress may be a pre-requisite for aflatoxin production  transcription factor AtfB regulates the aflatoxin biosynthesis genes fas-1, ver-1, and omtA as well as antioxidant genes encoding for catalase and SOP , reducing the accumulation of oxidative compounds, and regulating glutathione redox in A. flavus mycelia (A. flavus mycelia and reduce aflatoxin biosynthesis. This hypothesis seems plausible since the expression of the maize Ethylene Responsive Factor 1 (ZmERF1), a key transcription factor involved in ethylene and JA signaling, was found to be higher in the immature kernel tissues of the resistant maize inbred TZAR101 , the metabolic precursor to ethylene, was capable of reducing the expression of  mycelia . Therefo TZAR101  in compaculation .A. flavus tend to accumulate antioxidant enzymes, such as peroxidase and SOP, and tend to be more resistant to drought and heat stress than varieties susceptible to A. flavus  followed by increased catalase activity beginning at three DPI in maize kernels (Previous research has shown that maize varieties resistant to . flavus . Given t kernels , a combi kernels .A. flavus infection and mycotoxin biosynthesis are critical areas of research for the mitigation of aflatoxin contamination in maize. Maize resistance to A. flavus is a complex, quantitative trait which is the culmination of the interaction of numerous resistance-associated proteins and antioxidant enzymes which have been the subject of more than 50 years of rigorous research. Solutions to the problem of aflatoxin contamination of crops, particularly maize, have been elusive given the high level of environmental influence on the interaction and the lack of stable resistance in maize germplasm across multiple environments. By better understanding the role of oxidative stress and its remediation by the host and the pathogen, additional tools will be made available to counter the threat aflatoxin poses to food safety and security and further enhance the knowledge of cross-kingdom interactions which may be applied to other mycotoxin producing pathogens in various agricultural commodities.Determining the role of oxidative stress in the regulation of aflatoxin biosynthesis as well as the role of host defenses against both The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The development of surgical techniques differs greatly from that of new drugs. Surgical procedures often become widely used and accepted without any evidence from RCTs to show their superiority or non-inferiority to alternative medical treatments.This phenomenon, together with varying waiting-times for surgeries and patient expectations can place a number of constraints on the design of surgical trials, which impact greatly on the planned analysis of such studies. Often, these factors are working against regulators' increasing awareness of the need to thoroughly assess the effectiveness of surgical procedure.This presentation discusses some of these challenges and demonstrates how these were approached in CSAW, a trial assessing the effectiveness of sub-acromial decompression surgery for shoulder pain.There are conflicting demands on trial design and planned analysis. The most evident of these is that clinical researchers are interested in the effect of the treatment from the intervention (i.e. the surgery), whereas conventional statistical techniques place outcome assessments at specific time-points from randomisation. Differences between both strategies are particularly pronounced in trials measuring short-term outcomes or comparing surgical treatments with potentially long waiting-times and non-surgical interventions. There is tension between the need to accurately reflect the effects of a major intervention compared to need to account for any other effects, including the passage of time. Both conflicting viewpoints have advantages and limitations. This paper considers the decision-making process leading to a trial design and analysis plan that considers a compromise between appropriate trials methodology, patient concerns and expectations, as well as surgeon objectives.


Medical imaging provides information valuable in diagnosis, planning, and control of therapies. In this paper, we develop a method that uses a specific type of imaging—the magnetic resonance thermometry—to identify accurate and computationally efficient site and patient-specific computer models for thermal therapies, such as focused ultrasound surgery, hyperthermia, and thermally triggered targeted drug delivery. The developed method uses a sequence of acquired MR thermometry images to identify a treatment model describing the deposition and dissipation of thermal energy in tissues. The proper orthogonal decomposition of thermal images is first used to identify a set of empirical eigenfunctions, which captures spatial correlations in the thermal response of tissues. Using the reduced subset of eigenfunction as a functional basis, low-dimensional thermal response and the ultrasound specific absorption rate models are then identified. Once identified, the treatment models can be used to plan, optimize, and control the treatment. The developed approach is validated experimentally using the results of MR thermal imaging of a tissue phantom during focused ultrasound sonication. The validation demonstrates that our approach produces accurate low-dimensional treatment models and provides a convenient tool for balancing the accuracy of model predictions and the computational complexity of the treatment models. In ultrasound (US) thermal therapies, the goal is to selectively heat the treatment target without excessively elevating the temperature in healthy tissues intervening in the path of transmitted US energy and surrounding the target. In recent years, temperature measurements obtained by MR thermometry has played an increasingly important role in planning and control of thermal therapies. A number of techniques have been developed to obtain MR temperature maps, with water proton resonance frequency (PRF) method being the most widely used in practice In this paper, we develop and validate methods that avoid pointwise utilization of imaging data, leading to a highly efficient compression of MR thermometry images and the identification of a low-dimensional dynamic treatment models. Fundamentally, a low-dimensional representation of MR thermal images is possible because the image voxels are spatially correlated, reflecting the dependence of temperature distribution in the treatment target on the specific absorption rate (SAR) of the ultrasound and the heat dissipation by conduction and convection. Furthermore, a time series of MR thermal images are correlated temporally because of the causal dependance of temperatures on the heating history and temperatures at the preceding times. In the developed approach, we apply proper orthogonal decomposition (POD) to identify orthonormal basis functions Approximate each image acquired during therapy in the reduced set of basis functions. This step may be viewed as a real time image compression, which exploits spatial correlations of image voxels, as well as the image filtering step since high-frequency spatial valuations, which usually correspond to MRI measurement noise, are removed.Identify the ultrasound specific absorption rate and the dynamic model that captures temporal correlation in the series of images and provides a prediction the evolution of the ultrasound treatment.in vitro MRI tissue phantom heated by a focused ultrasound transducer. The results show that the identified low-dimensional models predict the SAR and the temperature response of the target with the expected accuracy, which can be controlled by selecting the order The developed approach is validated experimentally, using an known infinite-dimensional dynamic system, which makes it a popular approach in a variety of applications, including image processing The proper orthogonal decomposition is a technique often used to extract a set of basis functions for an approximate, modal-like representation of an infinite-dimensional, distributed parameter system (DPS). It was previously shown unknown infinite-dimensional model, characterizing the heat transport in the target due to noninvasive ultrasound heating. The following brief outline gives the exposition to the POD method in the context of our objectives. Further theoretical and algorithmic details are available in In this paper, we use a time series of images to identify an orthonormal basis of an Consider a set of MR thermal images (snapshots) The optimization problem (1) is difficult to solve in the general case. The problem is simplified if, following Assuming that all eigenvalues are ordered , can be adequately approximated by model (14). Consider, for example, the following popular Pennes' bioheat transfer equation (BHTE) Matrix The problem is to identify In order to decouple the problems of identifying The specific absorption rate of an ultrasound transducer depends on tissue properties and other variable factors, which can change with treatment site and from patient to patient. At the same time, the knowledge of the patient- and site-specific SAR is critically important for treatment safety and its precise control.Within the developed approach, the identification of the SAR is accomplished easily. Once the system matrix et al.The developed approach to identifying low-dimensional models of ultrasound therapies was tested experimentally using MR measurements of the thermal response of a tissue phantom to focused ultrasound (FUS) sonication. The cubic Temperature increase, Following the PRF shift method A step increase from zero to either 3.5, 4.8 or 6.5W of total electrical power (direct minus reflected) was applied to the FUS transducer while keeping all other experimental conditions the same. The phantom was allowed to reach thermal equilibrium before changing the power input.training (estimation) dataset to identify the reduced-order basis, and the corresponding models of the thermal response and the ultrasound SAR. The validation datasets, consisting of the images collected during the experiments with the other two power levels, were used to assess the adequacy of the identified basis functions and the accuracy of the temperature predictions obtained with the identified ultrasound treatment model.MRI thermal images collected for the case of 6.5W of applied electrical power were used as a A total of 499 MR thermal images were acquired to characterize temperature evolution during each power step test. The estimation dataset included images collected when the power was kept constant at 6.5W , the model-generated The developed approach was shown to be effective in identifying low-dimension but accurate models of ultrasound thermal therapies. At a pre-treatment stage, a set of MR thermometry images, characterizing the response of the target and the surrounding normal tissue to thermal excitation, is collected and then used to identify the reduced POD basis, which capture spatial correlations in images. A simple criterion for selecting an appropriate number of basis functions is provided which allows a user to balance the computational complexity of a predictive treatment model with its computational complexity. The selected reduced basis is then used to parsimoniously approximate newly acquired images, thus minimizing storage and data traffic. As an additional benefit, image approximation in the reduced basis filters high-frequency spatial noises in MR images.The SAR and thermal response models identified following the developed approach are patient- and site-specific and can be used as predictive models in real time  and off line (such as treatment planning) applications. The identification procedures are well suited to perform continuous re-identification of treatment models during the therapy. Such intra-treatment adaptability helps to mitigate the effect of changing tissue properties (such an US absorption) and blood perfusion, caused by elevated temperatures, on the accuracy of model predictions, which is particularly important when the identified low-dimensional models are used to efficiently implement model-based, optimizing treatment controllers that utilize images in the feedback. A family of related results used to identify continuous-time treatment models is described in reference in vitro MR experiments with a tissue phantom heated by a focused ultrasound transducer. The experimental results indicate that the SAR and thermal response during the treatment can be accurately predicted by the identified projection models with only two states. The low-dimensionality of the identified models substantially minimizes computational requirements of implementing a model-based treatment control system and communication traffic between the MRI scanner and the treatment controller.The developed methods were validated during Though our emphasis is on thermal therapies, the developed approach has a broader applicability in image-based identification and image-guided control of therapies. After straightforward modifications, this approach can be used with images acquired in multiple planes and with three-dimensional MR measurements.


Cafeteria roenbergensis virus, shares several genes with large eukaryotic self-replicating transposon of the Polinton (Maverick) family, and it has been proposed that the polintons evolved from a Mavirus-like ancestor.Recent advances of genomics and metagenomics reveal remarkable diversity of viruses and other selfish genetic elements. In particular, giant viruses have been shown to possess their own mobilomes that include virophages, small viruses that parasitize on giant viruses of the Mimiviridae family, and transpovirons, distinct linear plasmids. One of the virophages known as the Mavirus, a parasite of the giant Tetrahymena transposable element Tlr1, transpovirons, adenoviruses, and some bacteriophages form a network of evolutionary relationships that is held together by overlapping sets of shared genes and appears to represent a distinct module in the vast total network of viruses and mobile elements.We performed a comprehensive phylogenomic analysis of the available genomes of virophages and traced the evolutionary connections between the virophages and other selfish genetic elements. The comparison of the gene composition and genome organization of the virophages reveals 6 conserved, core genes that are organized in partially conserved arrays. Phylogenetic analysis of those core virophage genes, for which a sufficient diversity of homologs outside the virophages was detected, including the maturation protease and the packaging ATPase, supports the monophyly of the virophages. The results of this analysis appear incompatible with the origin of polintons from a Mavirus-like agent but rather suggest that Mavirus evolved through recombination between a polinton and an unknownvirus. Altogether, virophages, polintons, a distinct The results of the phylogenomic analysis of the virophages and related genetic elements are compatible with the concept of network-like evolution of the virus world and emphasize multiple evolutionary connections between bona fide viruses and other classes of capsid-less mobile elements. Cafeteria roenbergensis virus (CroV), a distant relative of the mimiviruses [The rapid advances of genomics and metagenomics lead not only to the rapid growth of sequence databases but to discovery of fundamentally novel types of genetic elements. The discovery and characterization of giant viruses that infect unicellular eukaryotes, in particular members of the family Mimiviridae infecting amoeba, over the last decade revealed a remarkable new class of agents that are typical viruses by structure and reproduction strategy but exceed many parasitic bacteria in size and genomic complexity -6. Much iviruses ,11. The iviruses  but accoiviruses . Very reiviruses . Four coiviruses .The three well-characterized virophages possess small isocahedral virions and genomes of 20 to 25 kilobase encoding 21 to 26 proteins each. Although the virophages are similar in genome size and structure and are generally construed as related, only a minority of the virophage genes are homologous. The rest of the genes show diverse phylogenetic affinities suggestive of chimeric origins of the virophages ,11,12.Trichomonas vaginalis, have long been considered ‘virus-like’ transposons because of their large size (20 kb and larger) and the presence of several genes that are common in viruses but not in other transposable elements such as B family DNA polymerase (PolB), packaging ATPase (ATPase) and protease (PRO) [Analysis of the Mavirus genome  resultedse (PRO) -18. The se (PRO) .In addition to the virophages, the giant viruses host several other groups of mobile elements. These include self-splicing introns, inteins, putative bacterial-type transposons and the most recently discovered novel linear plasmids named transpovirons . The traWe sought to decipher the evolutionary relationships between the three known virophages, the Polintons, transpovirons and possibly other genetic elements and viruses. We come up with a complex network of evolutionary relationships that connect many of these diverse elements through overlapping sets of homologous genes.To our knowledge, the evolutionary relationships between the virophages so far have not been analyzed in a comprehensive manner. Therefore we performed an exhaustive genomic comparison of the three well-characterized virophages that involved detailed sequence analysis for all predicted virophage proteins (see Methods for details); at this stage, the 5 new virophage genome sequences  were notThe virophage Zn-ribbon is a distinct version of this module that is shared by the virophages and several other groups of mobile elements (see below). In the Sputnik virophage the Zn-ribbon is a stand-alone protein whereas in Mavirus and OLV it is fused to a GIY-YIG endonuclease (GIY), a domain architecture that was detected also in environmental homologs. Conceivably, the ZnR-nuclease fusion is the ancestral version of this protein, with the nuclease lost in the Sputnik lineage. The ZnR domain is too small for reliable phylogenetic analysis  encode a Superfamily 3 helicase (S3H) domain that in Sputnik, OLV and YSLV1 is fused to the N-terminal domain belonging to a distinct family of polymerases-primases (TVpol) homologous to the bacterial DNA polymerase I . By contSputnik and OLV share two proteins (or domains) that are missing in Mavirus including the primase domain discussed above and an uncharacterized protein V21/OLV5  that are missing in Sputnik. One of these is the GIY-YIG endonuclease domain that is encoded in two genes in OLV and in a single gene in the Mavirus Figure  and is fSputnik and Mavirus exclusively share only one pair of homologous genes that encode a catalytic subunit of integrase with homologs in numerous bacterial and eukaryotic transposons. The Sputnik integrase appears to share a common ancestry with bacteriophage integrases , whereasThe conservation and the demonstrable monophyly of the two capsid protein genes and the key proteins involved in the virion maturation, the protease and the packaging ATPase, imply that the virophages evolved from a common ancestor that was a bona fide virus. In addition to the genes that are conserved in all virophages, the parsimony principle combined with the phylogenetic tree topologies dictates that those genes that are shared by the Mavirus and either Sputnik or OLV are tentatively assigned to the ancestral virophage as well. In practice, there seems to be only one such gene, the GIY-YIG endonuclease containing a ZnR domain Figure .Beyond the conclusion on the existence of an ancestral virophage, the comparative analysis of the 3 virophage genomes, and in particular the complex history of the helicase-primase gene (see above), seem to be compatible with either of two distinct evolutionary scenarios Figure . Taking The second evolutionary scenario postulates that the Sputnik-OLV genome architecture including the primase-helicase fusion gene is ancestral to virophages whereas the PolB and INT genes were acquired by the Mavirus lineage along with the loss of the TVpol domain; under this scenario, the displacement of the primase-helicase with a distinct helicase domain occurred in the Mavirus lineage Figure B. This sRegardless of the exact evolutionary scenario, the virophages clearly combine genes from several different sources as noticed in the original report on Sputnik  Table 11. ModulaRemarkably, each of the virophages possesses genes that are closely related to homologs from their specific giant virus hosts Table . MoreoveThe Polintons show notable variability of the gene repertoire but possess a conserved core of 4 genes that consists of PolB, integrase, a C5-family protease and a packaging ATPase Figure . All theThe phylogenetic tree topologies of the virophage genes show much uncertainty, presumably caused by the small size of the conserved domains and their high sequence divergence that probably reflects high and non-uniform evolutionary rates in virophages, other viruses and polintons. Nevertheless, the key observation in the phylogenetic analysis of the genes that are shared by Mavirus with polintons seems to be that Mavirus  does not cluster with the polintons as a group but rather falls inside the polinton subtree. This topology of the phylogenetic trees appears incompatible with the origin of the polintons from a Mavirus-like ancestor as previously proposed . InsteadUnder each of the two distinct scenarios shown in Figure Transpovirons represent a novel class of mobile elements, apparently linear plasmids that so far have been identified only in association with mimiviruses . RemarkaT. thermophila[Megavirus chiliensis[Guillardia theta , can beiviruses ,32. The Another important outcome of this analysis is the demonstration of multiple connections between bona fide viruses that encode capsid proteins and form infectious viruses and non-viral mobile elements such as transposons. It appears that viruses evolved from non-viral genetic elements and vice versa on more than one occasion even within this relatively small module of the virus evolution networks. These findings imply that capsid-centric concepts of virus evolution ,34 captuhttp://www.ncbi.nlm.nih.gov/Web/Newsltr/Spring04/blastlab.html); a representative (the longest) sequence from each cluster was taken. Protein sequences were aligned using MUSCLE [The protein sequences were extracted from the RefSeq database  . The nong MUSCLE ; gapped g MUSCLE . A prelig MUSCLE ; the besg MUSCLE ; and theg MUSCLE , with thThe branch support values were expressed in Expected-Likelihood Weights (ELW). For S3H helicase, alternative tree topologies were tested with TreeFinder using the approximately unbiased (AU) test . In addiThe authors declare that they have no competing interests.EVK initiated and designed the study; NY and EVK collected and analyzed data; EVK and DR wrote the manuscript that was read, edited and approved by all authors.Multiple alignments of conserved virophage proteins/domains: MCP, mCP, GIY, ZnR, and Mavirus peptidase mv19.Click here for filePrimase-helicase topology testing.Click here for fileMultiple sequence alignments and phylogenetic trees computed using TreeFinder and PhyML (Newic format).Click here for file


Despite numerous advances in health care practices, cardiovascular disease still remains the leading cause of morbidity and mortality worldwide. Perhaps the most important consequence of cardiovascular disease is the interruption of blood flow to organs such as the heart and brain, resulting in the clinical presentation of a heart attack or stroke. As such, the regulation of vascular tone and the maintenance of vascular patency are vital for the preservation of cardiovascular health. Central to this process is the vascular endothelium. The endothelium is vital for the regulation of vascular tone and the maintenance of vascular homeostasis, as it releases factors such as nitric oxide, hydrogen sulfide, endothelial-dependent hyperpolarizing factor, and prostacyclin that induce vasodilatation and keep the vasculature free of obstructions . TherefoThis special issue of highlights the latest findings related to the role of endothelial dysfunction in cardiovascular disease and comprises five review articles and one original report. The first three articles share the common theme of endothelial dysfunction in diabetes. Each paper offers unique insights into the mechanisms underlying diabetes-induced changes in endothelial biology and highlights specific pathways that could be novel therapeutic targets.Endothelial dysfunction and diabetes: effects on angiogenesis, vascular remodeling, and wound healing,” discusses how ROS and deficiencies in NO bioavailability contribute to altered angiogenesis in diabetes. Therapeutic management of angiogenesis in diabetes is also discussed.The review article in this series by S. D. Funk et al. discusses the mechanisms that contribute to the development of atherosclerosis in diabetics. This paper highlights the central role of hyperglycemia in promoting endothelial dysfunction and discusses how cellular and animal studies have translated to clinical development of therapeutics targeting endothelial dysfunction in diabetes. The next paper by A. Sharma et al. highlights the importance of alterations in endothelium-derived nitric oxide (EDNO) production in the pathogenesis of vascular complications in both type 1 and type 2 diabetes. The interrelationship between reactive oxygen species (ROS) and EDNO is described, with emphasis on the cellular and molecular mechanisms that give rise to altered NO bioavailability in diabetes. In their paper, G. K. Kolluru et al. emphasize the importance of angiogenesis in diabetes and chronicle studies that have yielded mechanistic insights into the development of endothelial dysfunction in diabetes. This comprehensive paper, entitled “In another review article in this series, M. Murakami provides a comprehensive overview of the mechanisms contributing to the active maintenance of the vasculature. The role of growth factors and cytokines in mediating endothelial and mural cell maintenance is described, with an analysis of how genetic alterations in these pathways give rise to blood vessel abnormalities and susceptibility to vascular disease. The molecular and cellular mechanisms leading to restenosis and the potential for new therapeutics targeting vascular smooth muscle cell (VSMC) proliferation are the topic of a review article in this series. L. Denes et al. highlight the importance of VSMC phenotype switching and proliferation in vascular disease and discuss the clinical diagnosis and management of restenosis.Bradykinin type-2 receptor expression correlates with age and is subjected to transcriptional regulation.” This study demonstrates that bradykinin type-2 receptor (BK-2R) mRNA is positively correlated with age, but is significantly reduced in human patients with idiopathic dilated cardiomyopathy (IDC) compared with healthy individuals. The study further examines the relationship between polymorphisms in the BK-2R promoter and the presence of IDC and coronary heart disease.The original report by I. Liesmaa et al. is entitled “


Transthoracic echocardiography (TTE) is a reliable, noninvasive imaging method that is useful in the evaluation of cardiovascular thrombosis. We conducted a retrospective study of all the echocardiograms from patients in the postoperative care unit to assess the role of TTE in thrombus identification in the left ventricle.This retrospective database evaluation included all echocardiograms during a 14-month period. The echocardiographic examination protocol included the subcostal four-chamber view, the apical four-chamber view, the apical two-chamber view and the parasternal view, along the long and short axes in both spontaneously and mechanically ventilated patients. All echocardiograms were obtained within the 48 hours immediately following surgery.In total, 160 postoperative echocardiograms were obtained from 160 patients and resulted in the detection of five cases of left ventricular thrombosis. Subgroup analysis showed that 21 and 35 of the 160 patients examined had either dilated or ischemic cardiomyopathy, respectively. In these patients, preoperative echocardiograms had been obtained recently prior to surgery and were negative for left ventricular thrombus. In three of 35 patients with ischemic cardiomyopathy and two of 21 patients with dilated cardiomyopathy, thrombus was identified in the left ventricle. The thrombi were mobile, uncalcified and pedunculated and were located in the apex of the left ventricle. In addition, no clinical consequences of the left ventricular thrombi were recorded.Low-flow conditions in heart chambers due to ischemic or dilated cardiomyopathy in conjunction with the hypercoagulability caused by perioperative prothrombotic factors may lead to thrombotic events in the left ventricle. Echocardiography can significantly alter the management of up to 46% of critically ill patients -4. TTE aet al.  have advet al. .In addition, focused ultrasonography has emerged as an important and noninvasive bedside diagnostic tool for the emergency room physician that facilitates the early detection of potentially reversible and time-dependent conditions. Currently, the two primary indications for TTE are the diagnosis of pericardial tamponade and the confirmation or refutation of pulseless electrical activity .In this retrospective study, we report our experience using TTE for thrombus identification in the left ventricle in the setting of a postoperative care unit.We conducted a retrospective study of all echocardiograms from patients in the postoperative care unit to assess the role of TTE in thrombus identification in the left ventricle during the postoperative period. The postoperative care unit receives both elective and emergency admissions from a wide range of surgical specialties, including major vascular, thoracic, abdominal and orthopedic surgery.Because of the retrospective design of the study, formal research ethics committee approval and patients' written informed consent for publication of this manuscript and accompanying images were deemed unnecessary after consultation with the local ethics committee.A retrospective database evaluation was undertaken for all echocardiograms obtained during a selected 14-month period. All echocardiograms examined were part of a specific echocardiographic protocol that is performed as a local standard of care for typical postoperative indications, including evaluation of left and right ventricular function, hypotension, pulmonary edema, diagnosis of pericardial effusion, suspected valvular disease and refractory hypoxia.A competent anesthetist consultant who was undertaking European Society of Echocardiography accreditation performed the TTE studies. Hand-carried ultrasound examinations were conducted using a 2- to 5-MHz phased array transducer on a portable ultrasound unit .The echocardiographic examination protocol included visualization of the subcostal four-chamber view, the apical four-chamber view, the apical two-chamber view and the parasternal long-axis and short-axis views in either spontaneously breathing or mechanically ventilated patients.All results were digitally archived to permit peer review, and consultant cardiologists reviewed ambiguous results.All echocardiography was performed within the 48 hours immediately following surgery. During the 14-month period, 160 postoperative echocardiograms were obtained from 160 patients  who had a median age of 67 years and who ranged in age from 20 to 89 years. In total, 125 of the 160 patients were spontaneously breathing, whereas the remaining 35 patients were under mechanical ventilation. The echocardiograms were obtained from patients who had undergone the following operations: major thoracic (10 patients), orthopedic (40 patients), vascular (35 patients) or abdominal (75 patients). In 45 of the 75 abdominal operations, the incision was in or extended into the upper abdomen.Five of 160 patients were found to have masses consistent with thrombi in the left ventricle. All of the thrombi were detected in the acute setting when TTE was performed and, specifically, were identified in the left ventricular apex. Table Clear visualization of the thrombi was observed in the apical four-chamber view, the apical two-chamber view and the parasternal short-axis view. In one of the cases, the thrombus was also visualized in the parasternal long-axis view Figure . FurtherSubgroup analysis showed that 21 and 35 of 160 patients examined had dilated and ischemic cardiomyopathy, respectively. In these patients, preoperative echocardiograms had been obtained recently prior to surgery and were negative for left ventricular thrombus. In 3 (8.5%) of 35 patients with ischemic cardiomyopathy and 2 (9.52%) of 21 patients with dilated cardiomyopathy, thrombus was identified in the left ventricle  and frequently have other thrombotic risk factors  ,13.Patients at risk for the development of a left ventricular thrombus are readily identified with echocardiography. Thrombi generally involve the apex of the left ventricle, most often in the presence of akinesis or dyskinesis. Although myocardial infarction is the most common predisposing cause of left ventricular thrombi, left ventricular thrombi can develop in any situation in which low flow occurs .In our study, all of the left ventricular thrombi were detected in the acute setting when TTE was performed immediately after surgery for hemodynamic monitoring purposes. In the left ventricle, thrombi were located only at the cardiac apex. Ischemic cardiomyopathy in three patients and dilated cardiomyopathy in two patients may have contributed to this thrombus formation . AlthougAlthough surgical patients may have absolute contraindications for anticoagulant therapy immediately after surgery, it is not well known whether the nature, quality and presence of thrombus represent an absolute indication for full-dose anticoagulant treatment. In our cases, TTE revealed an uncalcified, fresh and extensively mobile structure in the apex of the left ventricle. Therefore, full-dose anticoagulant therapy was considered indispensable in avoiding the consequences of thrombus dislodgement into the bloodstream.The sensitivity of TTE for detecting left ventricular thrombosis ranges between 92% and 95%, with specificity of 86% to 88% . On the In TTE, large, protruding and highly mobile thrombi are readily seen from the apical window, while laminar thrombi that do not protrude into the cavity are likely to be missed . Poor imIn our study, thrombi in the left ventricle were mainly identified in the apical four-chamber view and by scanning the apex in the short-axis parasternal view. Using the long-axis parasternal view, only one case of left ventricle thrombosis was recognized. From this view, it was not always possible to visualize the left ventricular apex. Indeed, only when the transducer is moved to a lower interspace is the left ventricular apex included in the field . Among oIn addition, left ventricular thrombi could not be seen using the subcostal view. In the first, second and third patients, the main reasons were technical limitations related to foreshortening and to the inability to visualize the left ventricular apex from the subcostal view because of the position of the transducer relative to the cardiac apex ,23. In tAlthough this study is retrospectively designed, our observations offer important information in an otherwise unknown topic such as that of left ventricular thrombosis in surgical patients; hence, the hypothesis of the high rates of left ventricular thrombi in surgical patients with either ischemic or dilated cardiomyopathy remains to be confirmed in a prospective study.There is clear value in using TTE imaging of the heart for the purpose of cardiovascular evaluation and optimization in the postoperative period. This report demonstrates that information gained from TTE imaging contributes to the correct diagnosis of cardiovascular thrombosis in patients in the acute postoperative setting who have either dilated or ischemic cardiomyopathy.• TTE can provide serendipitous information in critical care patients and could also facilitate the diagnosis of systemic or local disorders.• Low-flow conditions in the heart chambers due to ischemic or dilated cardiomyopathy in conjunction with the hypercoagulability caused by surgical trauma and/or cancer may lead to left ventricular thrombosis during the perioperative period. The thrombi formed were located in the left ventricular apex and were mainly depicted in the apical four- and two-chamber views as well as in the parasternal short-axis view.ICU: intensive care unit; TEE: transesophageal echocardiography; TTE: transthoracic echocardiography.The authors declare that they have no competing interests.TS conceived of the study and performed all the echocardiography. AA provided analysis of ultrasound imaging data and drafted the manuscript. MT reviewed and archived ultrasound data. FP provided expert echocardiographic consulting. GK participated in the design of the study.


Sir,We read with great interest the paper by The fine characterization of the ‘evolutionary biology’ of activating BRAF mutation in melanoma progression is clearly relevant as it correlates with a more aggressive course of disease  of further metastatization. The lack of further modifications in the BRAF mutational condition, once melanoma has metastasized, can possibly result from absent/limited intratumor heterogeneity for the mutation in the metastatic disease. Supporting this idea, single-cell clones generated from the short-term metastatic melanoma cultures Mel 195 and Mel 313 retained the heterozygous BRAFV600E mutated and homozygous wild-type genotypes of the parental metastatic cells, respectively (Prompted by the findings of tastases . It is nondition . Additiogenotype . These fectively .On the basis of