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Trees with weeping shoot architectures are valued for their beauty and serve as tremendous resources for understanding how plants regulate posture control. ThePrunus persica(peach) weeping phenotype, which has elliptical downward arching branches, is caused by a homozygous mutation in theWEEPgene. Until now, little was known about the function of WEEP protein despite its high conservation throughout Plantae. Here, we present the results of anatomical, biochemical, biomechanical, physiological, and molecular experiments that provide insight into WEEP function. Our data suggest that weeping peach does not have defects in branch structure. Rather, transcriptomes from the adaxial (upper) and abaxial (lower) sides of standard and weeping branch shoot tips revealed flipped expression patterns for genes associated with early auxin response, tissue patterning, cell elongation, and tension wood development.This suggests that WEEP promotes polar auxin transport toward the lower side during shoot gravitropic response, leading to cell elongation and tension wood development. In addition, weeping peach trees exhibited steeper root systems and faster root gravitropic response, just as barley and wheat with mutations in theirWEEPhomologEGT2. This suggests that the role of WEEP in regulating lateral organ angles and orientations during gravitropism may be conserved. Additionally, size-exclusion chromatography indicated that WEEP proteins self-oligomerize, like other SAM-domain proteins. This oligomerization may be required for WEEP to function in formation of protein complexes during auxin transport. Collectively, our results from weeping peach provide new insight into polar auxin transport mechanisms associated with gravitropism and lateral shoot and root orientation.

Supplementary Tables 1-3. Supplemental Table 1. Plasma pharmacokinetics of DMS612 metabolites. Supplemental Table 2. Pharmacodynamic assessment of !-H2AX detection in PBMCs. Mean values for !-H2AX foci per cell with DMS612 doses of 1.5, 3, 5, 7, 9 and 12 mg/m2 at the indicated time in hours (h). Supplemental Table 3. Pharmacodynamic assessment of !-H2AX detection in hair follicles. Mean values for cell with more than 4 !-H2AX foci per cell in plucked scalp hairs with DMS612 doses of 1.5, 3 and 9 mg/m2 at the indicated time in hours (h).

Supplementary Tables 1-3. Supplemental Table 1. Plasma pharmacokinetics of DMS612 metabolites. Supplemental Table 2. Pharmacodynamic assessment of !-H2AX detection in PBMCs. Mean values for !-H2AX foci per cell with DMS612 doses of 1.5, 3, 5, 7, 9 and 12 mg/m2 at the indicated time in hours (h). Supplemental Table 3. Pharmacodynamic assessment of !-H2AX detection in hair follicles. Mean values for cell with more than 4 !-H2AX foci per cell in plucked scalp hairs with DMS612 doses of 1.5, 3 and 9 mg/m2 at the indicated time in hours (h). Supplemental Figure 1. Pharmacokinetics of DMS612 Metabolites.

Purpose: DMS612 is a dimethane sulfonate analog with bifunctional alkylating activity and preferential cytotoxicity to human renal cell carcinoma (RCC) in the NCI-60 cell panel. This first-in-human phase I study aimed to determine dose-limiting toxicity (DLT), maximum tolerated dose (MTD), pharmacokinetics (PK), and pharmacodynamics (PD) of DMS612 administered by 10-minute intravenous infusion on days 1, 8, and 15 of an every-28-day schedule.Experimental Design: Patients with advanced solid malignancies were eligible. Enrollment followed a 3+3 design. PKs of DMS612 and metabolites were assessed by mass spectroscopy and PD by γ-H2AX immunofluorescence.Results: A total of 31 patients, including those with colorectal (11), RCC (4), cervical (2), and urothelial (1) cancers, were enrolled. Six dose levels were studied, from 1.5 mg/m2 to 12 mg/m2. DLTs of grade 4 neutropenia and prolonged grade 3 thrombocytopenia were observed at 12 mg/m2. The MTD was determined to be 9 mg/m2 with a single DLT of grade 4 thrombocytopenia in 1 of 12 patients. Two patients had a confirmed partial response at the 9 mg/m2 dose level, in renal (1) and cervical (1) cancer. DMS612 was rapidly converted into active metabolites. γ-H2AX immunofluorescence revealed dose-dependent DNA damage in both peripheral blood lymphocytes and scalp hairs.Conclusions: The MTD of DMS12 on days 1, 8, and 15 every 28 days was 9 mg/m2. DMS612 appears to be an alkylating agent with unique tissue specificities. Dose-dependent PD signals and two partial responses at the MTD support further evaluation of DMS612 in phase II trials. Clin Cancer Res; 21(4); 721–9. ©2014 AACR.

AbstractInternal hernias after Roux-en-Y gastric bypass are one of the most common complications in bariatric surgery leading to hospital readmissions with or without surgery. Due to the insidious and non-specific clinical presentation combined with difficult radiological identification, internal hernias remain a diagnostic and therapeutic challenge. Delay in diagnosis and treatment can lead to severe morbidity and mortality. We report the case of a patient presenting an unusual intestinal obstruction due to two simultaneous and distinct internals hernias affecting the Petersen’s defect and the foramen of Winslow 4 years after a laparoscopic gastric bypass. A 41-year-old female patient with a history of robotic laparoscopic Roux-en-Y gastric bypass presented with proximal small bowel obstruction symptoms and epigastric pain. Computed tomography showed two simultaneous internal hernias, one affecting the Petersen’s defect and the other affecting the foramen of Winslow with an incarcerated transverse colon. We performed an explorative laparoscopy to reduce both hernias and, after an assessment of bowel viability by indocyanine green angiography, we closed both defects with interrupted non-absorbable sutures to prevent recurrence. The follow-up of more than 1 year has been uneventful. In the cases of obstruction and hospital admission after gastric bypass, we suggest that patients undergo a computed tomography directly after the initial examination. Internal hernia diagnosis is often delicate and requires surgical exploration since bowel strangulation can lead to dramatic outcomes. The operation can be safely done laparoscopically, and all internal hernia defects should be repaired with non-absorbable sutures.

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Microbial processes are critical to the function of freshwater ecosystems, yet we still do not fully understand the factors that shape freshwater microbial communities. Furthermore, freshwater ecosystems are particularly susceptible to effects of environmental change, including influx of exogenous nutrients such as nitrogen and phosphorus. To evaluate the impact of nitrogen loading on the microbial community structure of shallow freshwater lakes, water samples collected from Lake Shenandoah (Virginia, USA) were incubated with two concentrations of either ammonium, nitrate, or urea as a nitrogen source. The potential impact of these nitrogen compounds on the bacterial community structure was assessed via 16S rRNA amplicon sequencing. At the phylum level, the dominant taxa in Lake Shenandoah were comprised of Actinobacteria and Proteobacteria, which were not affected by exposure to the various nitrogen treatments. Overall, there was not a significant shift in the diversity of the bacterial community of Lake Shenandoah with the addition of nitrogen sources, indicating this shallow system may be constrained by other environmental factors.

The installation of automatic data acquisition systems, together with the use of machine learning, allow obtaining useful information on the behaviour of dams. In this contribution, an example of application for a machine learning based predictive model is presented. Specifically, the level in a piezometer and its association with the reservoir level is studied for an embankment dam. The results show the model’s ability to identify changes in dam response by taking full advantage of the available monitoring data. The flexibility of the algorithm allows different types of variables to be analysed without the need to determine a priori which are the most influential loads or how they affect the target value. The model has been implemented in a software tool that includes additional functionalities, specific for the treatment and exploration of dam monitoring data. It can be applied to different dam types and response variables.

Supplemental Digital Content is available in the text., Rationale: Plaque rupture is the proximate cause of most myocardial infarctions and many strokes. However, the molecular mechanisms that precipitate plaque rupture are unknown. Objective: By applying proteomic and bioinformatic approaches in mouse models of protease-induced plaque rupture and in ruptured human plaques, we aimed to illuminate biochemical pathways through which proteolysis causes plaque rupture and identify substrates that are cleaved in ruptured plaques. Methods and Results: We performed shotgun proteomics analyses of aortas of transgenic mice with macrophage-specific overexpression of urokinase (SR-uPA+/0 mice) and of SR-uPA+/0 bone marrow transplant recipients, and we used bioinformatic tools to evaluate protein abundance and functional category enrichment in these aortas. In parallel, we performed shotgun proteomics and bioinformatics studies on extracts of ruptured and stable areas of freshly harvested human carotid plaques. We also applied a separate protein-analysis method (protein topography and migration analysis platform) to attempt to identify substrates and proteolytic fragments in mouse and human plaque extracts. Approximately 10% of extracted aortic proteins were reproducibly altered in SR-uPA+/0 aortas. Proteases, inflammatory signaling molecules, as well as proteins involved with cell adhesion, the cytoskeleton, and apoptosis, were increased. ECM (Extracellular matrix) proteins, including basement-membrane proteins, were decreased. Approximately 40% of proteins were altered in ruptured versus stable areas of human carotid plaques, including many of the same functional categories that were altered in SR-uPA+/0 aortas. Collagens were minimally altered in SR-uPA+/0 aortas and ruptured human plaques; however, several basement-membrane proteins were reduced in both SR-uPA+/0 aortas and ruptured human plaques. Protein topography and migration analysis platform did not detect robust increases in proteolytic fragments of ECM proteins in either setting. Conclusions: Parallel studies of SR-uPA+/0 mouse aortas and human plaques identify mechanisms that connect proteolysis with plaque rupture, including inflammation, basement-membrane protein loss, and apoptosis. Basement-membrane protein loss is a prominent feature of ruptured human plaques, suggesting a major role for basement-membrane proteins in maintaining plaque stability.

The CuA center is the initial electron acceptor in cytochrome c oxidase, and it consists of two copper ions bridged by two cysteines and ligated by two histidines, a methionine, and a carbonyl in the peptide backbone of a nearby glutamine. The two ligating histidines are of particular interest as they may influence the electronic and redox properties of the metal center. To test for the presence of reactive ligating histidines, a portion of cytochrome c oxidase from the bacteria Thermus thermophilus that contains the CuA site (the TtCuA protein) was treated with the chemical modifier diethyl pyrocarbonate (DEPC) and the reaction followed through UV–visible, circular dichroism, and electron paramagnetic resonance spectroscopies at pH 5.0–9.0. A mutant protein (H40A/H117A) with the non-ligating histidines removed was similarly tested. Introduction of an electron-withdrawing DEPC-modification onto the ligating histidine 157 of TtCuA increased the reduction potential by over 70 mV, as assessed by cyclic voltammetry. Results from both proteins indicate that DEPC reacts with one of the two ligating histidines, modification of a ligating histidine raises the reduction potential of the CuA site, and formation of the DEPC adduct is reversible at room temperature. The existence of the reactive ligating histidine suggests that this residue may play a role in modulating the electronic and redox properties of TtCuA through kinetically-controlled proton exchange with the solvent. Lack of reactivity by the metalloproteins Sco and azurin, both of which contain a mononuclear copper center, indicate that reactivity toward DEPC is not a characteristic of all ligating histidines.

As part of a comprehensive study to survey predators associated with hemlock woolly adelgid, Adelges tsugae Annand, 1928 in the Pacific Northwest (PNW), U.S.A. (Kohler et al. 2008), predators of balsam woolly adelgid, Adelges piceae (Ratzeburg, 1844) and Cooley spruce gall adelgid, Adelges cooleyi (Gillette, 1907) (all Hemiptera: Adelgidae) were sampled on nine dates from June 2005 to October 2006 on a small number of trees. Samples were collected every 6–8 weeks. Predators were collected from two A. piceae infested grand fir, Abies grandis (Douglas ex David Don) Lindley, and one A. cooleyi infested Engelmann spruce, Piceae engelmanii Parry ex Engelmann, on the Oregon State University campus in Corvallis, Oregon, and two A. cooleyi infested Douglas-fir, Pseudotsuga menziesii (Mirbel) Franco (all Pinaceae). One Douglas-fir was an ornamental tree in Corvallis, Oregon and the other was in a seed orchard on Whidbey Island, Washington. Adelges cooleyi predators were collected by striking an infested branch four times over a plastic tub with an area of 0.16 m2. Adelges piceae predators were collected by brushing adelgid wool on the bole of infested trees with a 2-inch wide paintbrush over a beat sheet. Predators were collected from the tub and beat sheet with an aspirator. Adult insects were killed in a sealed 9-dram plastic vial containing a small piece of Hot Shot No-Pest Insecticide Strip (Spectrum Brands Inc., Atlanta, Georgia). Immature insects were killed in KAAD mixture (10 parts 95% ethanol, 1 part kerosene, 2 parts glacial acetic acid, and 1 part dioxane) and preserved in 70% ethanol (Borrer et al. 1989). Adult specimens were identifi ed either by taxonomic specialists or by comparison to previously identified museum specimens. Voucher specimens were deposited in the Oregon State Arthropod Collection, Department of Zoology, Oregon State University, Corvallis, Oregon.

Purpose To evaluate in a proof-of-concept study the feasibility of Simultaneous Noncontrast Angiography and intraPlaque hemorrhage (SNAP) imaging as a clinical magnetic resonance angiography (MRA) technique for measuring carotid stenosis. There is a growing interest in detecting intraplaque hemorrhage (IPH) during the clinical management of carotid disease, yet luminal stenosis has remained indispensable during clinical decision-making. SNAP imaging has been proposed as a novel IPH imaging technique that provides carotid MRA with no added scan time. Flowing blood shows negative signal on SNAP because of phase-sensitive inversion recovery. Materials and Methods In all, 58 asymptomatic subjects with 16–79% stenosis on ultrasound were scanned at 3T by SNAP with 0.8 mm isotropic resolution and 16 cm longitudinal coverage. Two readers measured luminal stenosis of bilateral carotid arteries (n = 116) on minimum intensity projections of SNAP using the NASCET criteria. In the subset (48 arteries) with contrast-enhanced (CE) MRA available for comparison, luminal stenosis was also measured on maximum intensity projections of CE-MRA. Results Intraclass correlation coefficients (ICCs) with 95% confidence intervals were 0.94 (0.90–0.96) and 0.93 (0.88–0.96) for intra- and interreader agreement on stenosis measurements, respectively. Corresponding kappas for grading stenosis (0–29%, 30–69%, 70–99%, and 100%) were 0.79 (0.67–0.89) and 0.80 (0.68–0.90). Agreement between SNAP and CE-MRA was high (ICC: 0.95 [0.90–0.98]; kappa: 0.82 [0.71–0.93]). Conclusion As a dedicated IPH-imaging sequence, SNAP also provided carotid stenosis measurement that showed high intra- and interreader consistency and excellent agreement with CE-MRA. Further comparisons with digital subtraction angiography and other noninvasive techniques are warranted. Level of Evidence: 2 Technical Efficacy: Stage 2 J. Magn. Reson. Imaging 2017;46:1045–1052.

Nasopharyngeal carcinoma (NPC), an uncommon malignancy in Western Countries and Radiotherapy, remains an effective treatment. Its side effects are classified as either immediate or late; postradiation necrosis is as an important late side effect with a strong impact on the prognosis in patients with NPC. We report the case of 65-year-old Caucasian man presenting with a deep necrotic ulcer of the nasopharynx and osteoradionecrosis of the skull base that appeared 3 months after radiotherapy for nasopharyngeal carcinoma. Conservative treatment was applied with surgical management of the ulcer. Clinical and radiological outcomes are presented. Radiotherapy remains a good treatment option with varying degrees of side effects, in particular, postradiation necrosis and ulcer. Multiple options of treatment have been described. However, the surgical management could be indicated in cases of deep ulcer with life-threatening prognosis.

Adelges tsugae infested western hemlock trees were sampled periodically for 1 year at two locations in Oregon and Washington to compare the phenology and abundance of three associated predators (Leucopis argenticollis, Leucopis piniperda, and Laricobius nigrinus) and their host. On each sample date, two 3–10 cm long terminal twigs were collected from each tree and brought to the laboratory to count all life stages of A. tsugae and the three predators. Peak larval abundance of Leucopis spp. and La. nigrinus coincided with the presence of A. tsugae adults and eggs. Leucopis spp. larvae were present for a much longer period of time than were La. nigrinus larvae. Furthermore, Leucopis spp. larvae were present during both the progrediens and sistens egg stages, while La. nigrinus larvae were only present during the progrediens egg stage. Overall, we collected 2.3–3.5 times more Leucopis spp. of all life stages than La. nigrinus. These results support the continued study of Leucopis spp. from the Pacific Northwest as biological control agents for A. tsugae in the Eastern USA.

Background Intimal hyperplasia at the venous anastomosis of dialysis grafts causes early failure. We developed a sheep model of arteriovenous prosthetic grafts that fail rapidly due to intimal hyperplasia with histologic features nearly identical to human access grafts. A prominent feature of lesion development in this model is formation of luminal thrombus that becomes organized into stenosing lesions by macrophage and myofibroblast infiltration. To better understand this process, we examined the presence and activity of tissue factor (TF) in this system. This protein is the physiological initiator of coagulation in vivo and is known to contribute to development of intimal hyperplasia after vascular injury. Methods Expanded polytetrafluorethylene (ePTFE) grafts were placed between the carotid artery and external jugular vein in sheep. Grafts were examined for luminal TF activity using a novel ex vivo assay. In a separate series of grafts, immunohistochemistry was used to localize smooth muscle cells, monocytes, and TF protein. Results At 2 days, luminal TF activity already was higher in the venous and arterial end of the graft than in the adventitia. This high level of activity persisted at 8 weeks. TF activity was higher in the venous end of the grafts than in the arterial end at 2 and 8 weeks (40% and 47% increase, n = 5, n = 3, respectively, P

Las deformaciones principales de la pared anterior del torax son el pectus excavatum y el pectus carinatum. Las formas aisladas sin repercusion funcional grave son las mas frecuentes. La causa sugerida en la mayoria de los casos es una displasia de los cartilagos costales asociada a un desequilibrio de su crecimiento. Se pueden plantear tres principios terapeuticos. El primero tiene un planteamiento radical. Se trata de la reseccion de los cartilagos costales anormales o esternocondroplastias. La tecnica de condroplastia subpericondrica por una via de acceso horizontal minimamente invasivo permite tratar las formas complejas en pacientes adultos, y en menos ocasiones en la infancia. A esta tecnica se le asocia, en los casos de pectus excavatum, la colocacion de una ferula metalica retroesternal durante 6 meses. Las indicaciones de osteotomias esternales son actualmente infrecuentes. El segundo principio se basa en el modelado progresivo del torax durante el periodo de crecimiento toracico. La correccion se realiza con una placa de osteosintesis, que actua como un autentico tutor esternocostal temporal. Se puede introducir por una via minimamente invasiva con control toracoscopico. Esta tecnica es exclusiva del pectus excavatum simetrico en adolescentes. En las formas particularmente graves, un acceso subxifoideo minimo permite elevar el esternon y asegurar el paso de la placa. Se han descrito tecnicas recientes, como las campanas de succion, con resultados considerados prometedores, pero que no se han demostrado por el momento. En el pectus carinatum, una compresion externa mediante una ortesis proporciona buenos resultados demostrados. El tercer principio terapeutico es un tratamiento paliativo dirigido al relleno de la excavacion con una protesis de silicona. Su interes es la sencillez inmediata. Presenta el inconveniente de los riesgos de complicacion y de migracion del implante de silicona que se mantiene colocado varias decadas. En resumen, las indicaciones terapeuticas pueden decantarse por tecnicas quirurgicas diferentes, pero complementarias.

Practical application of silicon anodes for lithium-ion batteries has been mainly hindered because of their low electrical conductivity and large volume change (ca. 300%) occurring during the lithiation and delithiation processes. Thus, the surface engineering of active particles (material design) and the modification of electrode structure (electrode design) of silicon are necessary to alleviate these critical limiting factors. Silicon/carbon core–shell particles (Si@C, material design) are prepared by the thermal decomposition and subsequent three-dimensional (3D) electrode structures (electrode design) with a channel width of 15 μm are incorporated using the laser ablation process. The electrochemical characteristics of 3D Si@C used as the anode material for lithium-ion batteries are investigated to identify the effects of material and electrode design. By the introduction of a carbon coating and the laser structuring, an enhanced performance of Si anode materials exhibiting high specific capacity (>1200 mAh g −1 over 300 cycles), good rate capability (1170 mAh g −1 at 8 A g −1 ), and stable cycling is achieved. The morphology of the core–shell active material combined with 3D channel architecture can minimize the volume expansion by utilizing the void space during the repeated cycling.

Purpose: DMS612 is a dimethane sulfonate analog with bifunctional alkylating activity and preferential cytotoxicity to human renal cell carcinoma (RCC) in the NCI-60 cell panel. This first-in-human phase I study aimed to determine dose-limiting toxicity (DLT), maximum tolerated dose (MTD), pharmacokinetics (PK), and pharmacodynamics (PD) of DMS612 administered by 10-minute intravenous infusion on days 1, 8, and 15 of an every-28-day schedule. Experimental Design: Patients with advanced solid malignancies were eligible. Enrollment followed a 3+3 design. PKs of DMS612 and metabolites were assessed by mass spectroscopy and PD by γ-H2AX immunofluorescence. Results: A total of 31 patients, including those with colorectal (11), RCC (4), cervical (2), and urothelial (1) cancers, were enrolled. Six dose levels were studied, from 1.5 mg/m2 to 12 mg/m2. DLTs of grade 4 neutropenia and prolonged grade 3 thrombocytopenia were observed at 12 mg/m2. The MTD was determined to be 9 mg/m2 with a single DLT of grade 4 thrombocytopenia in 1 of 12 patients. Two patients had a confirmed partial response at the 9 mg/m2 dose level, in renal (1) and cervical (1) cancer. DMS612 was rapidly converted into active metabolites. γ-H2AX immunofluorescence revealed dose-dependent DNA damage in both peripheral blood lymphocytes and scalp hairs. Conclusions: The MTD of DMS12 on days 1, 8, and 15 every 28 days was 9 mg/m2. DMS612 appears to be an alkylating agent with unique tissue specificities. Dose-dependent PD signals and two partial responses at the MTD support further evaluation of DMS612 in phase II trials. Clin Cancer Res; 21(4); 721–9. ©2014 AACR.

β-Amylase3 (BAM3) is an enzyme that is essential for starch degradation in plant leaves and is also transcriptionally induced under cold stress. However, we recently reported that BAM3's enzymatic activity decreased in cold-stressed Arabidopsis leaves, although the activity of BAM1, a homologous leaf β-amylase, was largely unaffected. This decrease in BAM3 activity may relate to the accumulation of starch reported in cold-stressed plants. The aim of this study was to explore the disparity between BAM3 transcript and activity levels under cold stress, and we present evidence suggesting BAM3 is being inhibited by post-translational modification. A mechanism of enzyme inhibition was suggested by observing that BAM3 protein levels remained unchanged under cold stress. Cold stress induces nitric oxide (NO) signaling, one result being alteration of protein activity by nitrosylation or glutathionylation through agents such as S-nitrosoglutathione (GSNO). To test whether NO induction correlates with inhibition of BAM3 in vivo, plants were treated with sodium nitroprusside, which releases NO, and a decline in BAM3 but not BAM1 activity was again observed. Treatment of recombinant BAM3 and BAM1 with GSNO caused significant, dose-dependent inhibition of BAM3 activity while BAM1 was largely unaffected. Site-directed mutagenesis, anti-glutathione Western blots, and mass spectrometry were then used to determine that in vitro BAM3 inhibition was caused by glutathionylation at cysteine 433. In addition, we generated a BAM1 mutant resembling BAM3 that was sensitive to GSNO inhibition. These findings demonstrate a differential response of two BAM paralogs to the Cys-modifying reagent GSNO and provide a possible molecular basis for reduced BAM3 activity in cold-stressed plants.

Candida albicans is a natural component of the human microbiota but also an opportunistic pathogen that causes life-threatening infections. The human gastrointestinal tract is the main reservoir of C. albicans, from where systemic infections originate as a consequence of the disruption of the intestinal mucosal barrier. Recent studies provided convincing evidence that overgrowth of C. albicans and other related species in the gut is predominantly associated with chronic intestinal inflammatory bowel diseases. Here, we showed, for the first time, the antagonistic interkingdom interactions between C. albicans and common intestinal commensal bacteria. From a therapeutic perspective, administering a defined bacterial community, such as the one described here with anti-Candida activity, could provide potential therapeutic protection against gastrointestinal inflammatory diseases., Candida albicans is well known as a major human fungal pathogen, but it is also a permanent resident of healthy gastrointestinal tracts. Recent studies have shown that the human gut microbial metabolome represents an interesting source of bioactive molecules with a significant degree of chemical diversity. Some of these bioactive molecules may have useful antivirulence activities. For instance, intestinal bacterial species belonging to the Lachnospiraceae family were found to secrete molecules that attenuate Salmonella pathogenicity and repress the expression of virulence genes. Here, we have investigated whether the microbial gut metabolome (GM) contains molecules that might promote the commensal lifestyle and/or inhibit the expression of virulence of C. albicans in the intestine. We found that metabolites from human feces inhibited the growth of C. albicans and other opportunistic yeasts. A genetic screen in C. albicans suggested that TOR is the molecular target of the antifungal molecule(s) of the GM. In addition, we found that the GM metabolites inhibit both C. albicans hyphal growth and the invasion of human enterocytes. The antigrowth and antivirulence activities were partially recapitulated by secretions from Roseburia spp. and Bacteroides ovatus strains, respectively. This study demonstrates that the antimicrobial activity of the GM can be extended to a eukaryotic pathogen, C. albicans, illuminating the antagonistic interkingdom interactions between a fungus and intestinal commensal bacteria. IMPORTANCE Candida albicans is a natural component of the human microbiota but also an opportunistic pathogen that causes life-threatening infections. The human gastrointestinal tract is the main reservoir of C. albicans, from where systemic infections originate as a consequence of the disruption of the intestinal mucosal barrier. Recent studies provided convincing evidence that overgrowth of C. albicans and other related species in the gut is predominantly associated with chronic intestinal inflammatory bowel diseases. Here, we showed, for the first time, the antagonistic interkingdom interactions between C. albicans and common intestinal commensal bacteria. From a therapeutic perspective, administering a defined bacterial community, such as the one described here with anti-Candida activity, could provide potential therapeutic protection against gastrointestinal inflammatory diseases.

Target of rapamycin complex 1 (TORC1) is an essential regulator of metabolism in eukaryotic cells and in the fungal pathogen Candida albicans regulates morphogenesis and nitrogen acquisition. Gtr1 encodes a highly conserved GTPase that in Saccharomyces cerevisiae regulates nitrogen sensing and TORC1 activation. Here, we characterize the role of C. albicans GTR1 in TORC1 activation and compare it with the previously characterized GTPase Rhb1. A homozygous gtr1/gtr1 mutant exhibited impaired TORC1-mediated phosphorylation of ribosomal protein S6 and increased susceptibility to rapamycin. Overexpression of GTR1 impaired nitrogen starvation-induced filamentous growth, MEP2 expression, and growth in bovine serum albumin as the sole nitrogen source. Both GTR1 and RHB1 were shown to regulate genes involved in ribosome biogenesis, amino acid biosynthesis, and expression of biofilm growth-induced genes. The rhb1/rhb1 mutant exhibited a different pattern of expression of Sko1-regulated genes and increased susceptibility to Congo red and calcofluor white. The homozygous gtr1/gtr1 mutant exhibited enhanced flocculation phenotypes and, similar to the rhb1/rhb1 mutant, exhibited enhanced biofilm formation on plastic surfaces. In summary, Gtr1 and Rhb1 link nutrient sensing and biofilm formation and this connectivity may have evolved to enhance the competitiveness of C. albicans in niches where there is intense competition with other microbes for space and nutrients. IMPORTANCE Candida albicans is the major fungal pathogen of humans and is responsible for a wide range of infections, including life-threatening systemic infections in susceptible hosts. Target of rapamycin complex 1 (TORC1) is an essential regulator of metabolism in this fungus, and components of this complex are under increased investigation as targets for new antifungal drugs. The present study characterized the role of GTR1 , encoding a putative GTPase, in TORC1 activation. This study shows that GTR1 encodes a protein required for activation of TORC1 activity in response to amino acids and regulation of nitrogen starvation responses. GTR1 mutants show increased cell-cell adhesion and biofilm formation and increased expression of genes involved in these processes. This study demonstrates that starvation responses and biofilm formation are coregulated by GTR1 and suggests that these responses are linked to compete with the microbiome for space and nutrients.

PurposeDNA methylation is a well characterized epigenetic repressor of mRNA transcription in many plant and vertebrate systems. However, the mechanism of this repression is not fully understood. The process of synthesizing a strand of RNA from DNA, or transcription, is controlled by proteins that regulate RNA polymerase activity by binding to specific gene regulatory sequences. Cone-rod homeobox (CRX) is a well-characterized mammalian transcription factor that controls photoreceptor cell specific gene expression. While much is known about the functions and DNA binding specificity of CRX, less is known about how DNA methylation modulates CRX binding affinity to genomic cis-regulatory elements.MethodsWe used bisulfite pyrosequencing of human ocular tissues to measure DNA methylation levels of the regulatory regions of RHO, PDE6B, PAX6, and LINE. To describe the molecular mechanism of repression, we used molecular modeling to illustrate the effect of DNA methylation on human RHO regulatory sequences.ResultsIn this study, we demonstrate an inverse correlation between DNA methylation in regulatory regions adjacent to the human RHO and PDE6B genes and their subsequent transcription in human ocular tissues. Docking of CRX to our DNA models shows that CRX interacts with the grooves of these sequences, suggesting changes in groove structure could regulate binding. Molecular dynamics simulations of the RHO promoter and enhancer regions show changes in the flexibility and groove width upon epigenetic modification. Models also demonstrate that changes to the local dynamics of CRX binding sites within RHO regulatory sequences which may account for the repression of CRX dependent transcription.ConclusionCollectively, these data demonstrate epigenetic regulation of CRX binding sites in human retinal tissue and provide insight into the mechanism of this mode of epigenetic regulation to be tested in future experiments.

Resume Introduction Le pectus excavatum (PE) est une malformation congenitale frequente. La correction chirurgicale mini-invasive de Nuss (CCMI) consiste a relever par voie endoscopique le sternum par une plaque ou barre retrosternale. A partir d’une serie de 70 enfants, nous proposons d’en preciser les indications, notamment les limites. Materiel et methode Il s’agissait d’une etude retrospective et bicentrique. Soixante-dix enfants operes sont arrives en fin de croissance avec leur plaque retiree. La moyenne d’âge etait de 13,8 ans (6–19 ans). L’indication operatoire etait esthetique dans 66 cas (95 %). Soixante-trois ont ete operes selon la technique originale de Nuss. Sept patients ont necessite une voie d’abord complementaire sous-xiphoidienne. L’ablation de la plaque a ete realisee de 8 mois a 3 ans apres la pose. Resultats Le resultat esthetique a ete juge satisfaisant dans 64 cas (91 %) par les patients et 60 cas (85,7 %) par le chirurgien. Des complications majeures ont ete observees dans 6 cas (8,5 %) ayant necessite une reprise chirurgicale : 2 hemothorax, 2 sepsis parietaux (dont un apres ablation de la plaque), une allergie et une mobilisation de la plaque. Les complications mineures (46 cas [65 %]), precoces ou tardives, ont ete spontanement resolutives ou traitees medicalement. Discussion La faible rancon cicatricielle et la fiabilite du resultat obtenu sont en faveur de la diffusion de la technique de Nuss chez l’enfant et l’adolescent. Nos taux de complications etaient similaires aux donnees de la litterature (65 % mineures, 8,5 % majeures). Nous considerons que les patients presentant des antecedents de chirurgie cardiothoracique ou ceux presentant au scanner une distance sterno-vertebrale inferieure a 5 cm ou une rotation sternale superieure a 35° contre-indiquent une technique par voie endoscopique. Nous recommandons alors un controle sous- et retroxiphoidien ou une sternochondroplastie classique. Niveau de preuve Etude de cohorte descriptive, niveau 4.

Resume Une etude prospective multicentrique nationale a ete menee incluant les luxations congenitales de hanche diagnostiquees apres l’âge de 3 mois et impliquant outre la Societe francaise d’orthopedie pediatrique, l’Association francaise des pediatres ambulatoires et la Societe francophone d’imagerie pediatrique et prenatale. Elle revele une insuffisance de depistage clinique, evident sur le plan quantitatif et probablement qualitatif, requerant une campagne de communication et d’enseignement pour rappeler les bonnes pratiques. L’interet de l’echographie systematique n’est pas demontre. Des recommandations relatives au depistage de la LCH ont ete elaborees a la lumiere des resultats presentes dans cet article par les auteurs avec la Haute Autorite de sante. La realisation d’une etude nationale prospective randomisee multicentrique est indispensable, a laquelle il semble fondamental d’integrer les medecins generalistes.

Las deformidades de la pared toracica aparecen en la mayoria de los casos tras un exceso de crecimiento de los cartilagos costales que desplaza el esternon hacia dentro (pectus excavatum, lo mas frecuente) o, mas raramente, hacia fuera (pectus carinatum). En estos casos, la deformidad aparece y empeora progresivamente durante la infancia, con un pico maximo al inicio de la pubertad. La prescripcion de un tratamiento quirurgico u ortopedico se basa esencialmente en el perjuicio estetico, sobre todo en la adolescencia. Las manifestaciones cardiacas o respiratorias son raras, pero deben buscarse. Otras deformidades, claramente mas raras y a menudo congenitas, forman parte de sindromes complejos de dificil correccion.

Resume Contexte L’inconvenient majeur de la prise en charge des scolioses graves de survenue precoce (SGP) par la technique sans fusion avec distraction chirurgicale (tige de croissance conventionnelle) est la necessite de recourir a des chirurgies iteratives pendant l’enfance. La diffusion recente des tiges de croissance magnetiques (TCM) offre la possibilite d’eviter de telles chirurgies et par consequent, d’ameliorer la qualite de vie des patients. Hypothese Malgre le cout eleve des TCM et considerant leur impact potentiel sur la diminution du nombre de sejours hospitaliers, l’utilisation de ces tiges est susceptible de reduire la consommation de ressources medicales a long terme en comparaison des tiges de croissance conventionnelles (TCC). Materiel et methode Un modele d’evaluation medicoeconomique a ete construit afin d’evaluer et comparer le cout des deux strategies. Ce cout a ete estime a partir des probabilites de consommation de ressources medicales tirees d’une analyse de la litterature ainsi que des donnees d’une cohorte de patients avec SGP traites dans notre centre. Certaines ressources medicales ont egalement ete estimees a dire d’experts. Un horizon temporel de quatre ans a partir de la premiere implantation chirurgicale a ete retenu. Les couts ont ete evalues selon la perspective de l’assurance maladie francaise (en utilisant les tarifs de l’annee 2013) et ont ete actualises avec un taux annuel de 4 %. Des analyses de sensibilite ont ete conduites sur les differents parametres du modele afin d’evaluer sa robustesse. Resultats Sur un horizon temporel de quatre ans, les couts directs estimes pour les strategies TCC et TCM ont ete de 49 067 euros et 42 752 euros respectivement, soit un differentiel de cout de −6135 euros en faveur de la strategie TCM. Dans le premier cas, les couts etaient principalement representes par les sejours hospitaliers (83,9 %), alors que dans la strategie TCM, le cout de la tige a contribue pour 59,5 % du montant total. L’analyse de sensibilite univariee a permis d’identifier les parametres ayant la plus grande influence sur le differentiel de cout en particulier : le cout des sejours hospitaliers, le cout des TCM, et la frequence des distractions chirurgicales. Discussion La strategie TCM est une technique recente et prometteuse dans la prise en charge des SGP. En plus d’ameliorer la qualite de vie des patients, son utilisation dans le traitement des SGP est susceptible d’etre associee a une moindre consommation de ressources medicales. Niveau de preuve Niveau IV, etude medicoeconomique, etude retrospective.

Background Immune-mediated polyarthopathy (IMPA) is common in dogs, and is monitored by serial arthrocenteses. Hypothesis/Objectives Plasma C-reactive protein (CRP), interleukin-6 (IL-6), and CXCL8 (interleukin-8) would serve as noninvasive markers of joint inflammation in IMPA. Animals Nine client-owned dogs with idiopathic IMPA; 6 healthy controls. Methods Prospective study. Plasma CRP, IL-6, and CXCL8 were measured by ELISA at baseline, 2, and 4 weeks during treatment with prednisone at 50 mg/m2/day. Arthrocenteses, the canine brief pain inventory (CBPI), and accelerometry collars were used to assess joint inflammation, lameness, and mobility at all 3 time points. Results C-reactive protein concentrations were higher in IMPA dogs (median 91.1 μg/mL, range 76.7–195.0) compared with controls (median

Background: Neoantigens arise from DNA mutations in cancer cells and are important targets for T cell mediated anti-tumor immunity. NEO-PV-01 is a personal neoantigen vaccine of up to 20 peptides designed based on a patient’s neoantigen and HLA profile that is directed at inducing tumor-specific T cell responses to neoantigens. Here, we report relationships between baseline tumor characteristics, immune response and clinical outcome for NT-001, a Phase 1b study of NEO-PV-01 + adjuvant in combination with nivolumab in either first or later line therapy for patients with metastatic melanoma, bladder and non-small cell lung cancer (NCT02897765). This analysis is focused on the melanoma cohort. Methods: Serial blood and tumor biopsies were scheduled for collection: i) prior to treatment, ii) after 12 weeks of nivolumab monotherapy and iii) after completion of NEO-PV-01 vaccination. Baseline features and immune responses in tumor cells were characterized by immunohistochemistry for multiple immune and tumor markers, gene expression, whole exome and TCR sequencing. Antigen-specific responses by IFNγ ELISpot, intracellular cytokine staining, multi-parameter surface and functional phenotyping by FACS and the presence of cytolytic properties were monitored in serial peripheral blood samples. Durability of immune responses up to 104 weeks post start of treatment will be measured. Results: A cohort of 10 melanoma patients demonstrated CD4 and CD8 T cell responses against 56% of vaccine peptides that were detected primarily in the post-vaccination samples as measured by IFNγ ELISpot. Analysis of additional patients is ongoing. T cell responses were neoantigen-specific for most peptides tested (86%; 12/14). Vaccine-induced immune responses were durable in patients who reached the week 52 treatment timepoint. Most T cell responses were polyfunctional, as evident by secretion of multiple cytokines, exhibited a memory and effector memory phenotype and were cytolytic. Epitope spreading, defined as post-vaccination T cell responses to neoantigens not included in the vaccine, was observed in multiple melanoma patients analyzed and evaluated for association with post vaccine clinical responses. Further, multi-platform assessments of immune and molecular responses including gene expression and TCR repertoire analysis demonstrate extensive responses in patients continuing study past 52 weeks. Additional correlates of clinical outcomes with molecular and immunologic responses will be presented. Conclusions: Treatment with NEO-PV-01 + adjuvant in combination with nivolumab induced broad de novo neoantigen-specific immune responses in metastatic melanoma. Immune responses were specific and correlations with clinical outcomes will be discussed. Citation Format: Siwen Hu-Lieskovan, Patrick A. Ott, Aung Naing, Rana H. Besada, Samantha J. Gates, Victoria R. Kohler, Riley R. Curran, Meghan E. Bushway, Julian Scherer, Kristen N. Balogh, Tracey E. Sciuto, Ying S. Ting, Michael S. Rooney, Dewi Harjanto, Zhengping Huang, Yuting Huang, Yvonne Ware, April Lamb, Lisa D. Cleary, Melissa A. Moles, Richard B. Gaynor, Matthew J. Goldstein, Les H. Brail, Joel Greshock, Lakshmi Srinivasan. The personalized vaccine, NEO-PV-01 with anti-PD1, induces neoantigen-specific de novo immune responses in patients with advanced metastatic melanoma: Association with clinical outcomes [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 942.

Seagrasses provide a number of critical ecosystem services, including habitat for numerous species, sediment stabilization, and shoreline protection. Ariel photography is a useful tool to estimate the areal extent of seagrasses, but recent innovations in radiometrically calibrated sensors and algorithm development have allowed identification of benthic types and retrieval of absolute density. This study demonstrates the quantitative ability of a high spatial resolution (1 m) airborne hyperspectral sensor (3.2 nm bandwidth) in the complex coastal waters of Saint Joseph’s Bay (SJB). Several benthic types were distinguished, including submerged and floating aquatic vegetation, benthic red algae, bare sand, and optically deep water. A total of 23.6 km2 of benthic vegetation was detected, indicating no dramatic change in vegetation area over the past 30 years. SJB supported high seagrass density at depths shallower than 2 m with an average leaf area index of 2.0 ± 0.6 m2 m−2. Annual seagrass production in the bay was 13,570 t C year−1 and represented 41 % of total marine primary production. The effects of coarser spatial resolution were investigated and found to reduce biomass retrievals, underestimate productivity, and alter patch size statistics. Although data requirements for this approach are considerable, water column optical modeling may reduce the in situ requirements and facilitate the transition of this technique to routine monitoring efforts. The ability to quantify not just areal extent but also productivity of a seagrass meadow in optically complex coastal waters can provide information on the capacity of these environments to support marine food webs.

Fluorine-doped tin oxide (FTO) films with a thickness of about 3 micrometers were prepared by electron cyclotron resonance-metal organic chemical vapor deposition (ECR-MOCVD) under 800 W of microwave power, with tetra-methyl tin (TMT) as a tin precursor. The dome-shaped micro-patterned FTO layer was prepared on a copper current collector using a KrF excimer laser micromachining system for application as an anode for 3D lithium-ion batteries. Mild ECR plasma treatment at 600 W was carried out on the surface of the microstructured FTO anode, and the electrochemical characteristics were investigated with regard to the plasma treatment effects. The results show that physical properties such as the smooth and dense surface morphology and reduced surface oxygen functional groups of the plasma-treated samples enhanced the specific capacity, rate capability, and capacity fading. This was probably due to the reduction of side reactions, which may be closely related to the plasma treatment of the microstructured FTO layer. The ECR plasma treatment plays an important role in reducing the charging transfer resistance. In the experimental range studied, a higher specific capacity of 1425 mA h g−1 at a current density of 117 mA g−1 was observed, with capacity fading of 37.8% after 100 cycles for the plasma-treated microstructured FTO anode.

Resume La reconstruction diaphysaire apres resection d’une tumeur maligne demeure un challenge difficile. La technique de la membrane induite decrite par Masquelet, initialement indiquee dans les pertes de substance d’origine traumatique ou septique, commence a etre appliquee dans cette indication. Nous rapportons trois cas de lyse massive de la greffe apres reconstruction femorale, necessitant une ou plusieurs reprises chirurgicales chez deux garcons de trois et six ans et une fille de neuf ans. Cette complication pourrait s’expliquer par le delai important entre les deux temps operatoires, le greffon, les contraintes varisantes propres a la localisation femorale ou le defaut de stabilite de l’osteosynthese. L’application de cette technique dans les tumeurs osseuses a montre cependant des resultats preliminaires satisfaisants : c’est la localisation femorale qui semble specifiquement exposer a la lyse du greffon osseux. La recherche de l’origine de cette complication est fondamentale pour adapter la technique ou en modifier les indications.

Objective—The use of continuous-wave Doppler ultrasound to determine ankle brachial index (ABI) is the gold standard for diagnosing asymptomatic peripheral artery disease. This method is labor inte...

Resume Introduction L’osteome osteoide (OO) est une tumeur benigne, douloureuse dont le traitement reste chirurgical, dans l’objectif de realiser l’exerese du nidus. L’atteinte de la fosse acetabulaire est rare, mais pose le probleme de son accessibilite. Patients et methodes Nous presentons ici une serie de cinq patients atteints d’un osteome osteoide de la fosse acetabulaire pour lesquels a ete realise un forage resection osseux percutane (FROP) sous scanner. Resultats Pour ces cinq patients, les suites ont ete simples et l’evolution favorable avec une disparition immediate et durable des douleurs dans tous les cas. Discussion Les resultats de notre technique percutanee semblent aussi satisfaisants que les resultats des autres techniques percutanees, notamment en ce qui concerne la disparition des douleurs et la tolerance du patient vis-a-vis de la technique. Conclusion Le FROP sous controle tomodensitometrique a montre son efficacite therapeutique dans le cas d’une atteinte de la fosse acetabulaire par un osteome osteoide. Niveau de preuve IV.

The structure and phase formations of radio frequency magnetron sputtered lithium manganese oxide thin films (Li 1.1 Mn 1.9 O 4 ) under ambient air were studied. The influence of laser annealing and furnace annealing, respectively, on the bulk structure and surface phases was compared by using ex-situ X-ray diffraction and Raman analysis. Laser annealing technique formed a dominant (440)-reflection, furnace annealing led to both, (111)- and (440)-reflections within a cubic symmetry (S.G. Fd3m (227)). Additionally, in-situ Raman and in-situ X-ray diffraction were applied for online detection of phase transformation temperatures. In-situ X-ray diffraction measurements clearly identified the starting temperature for the (111)- and (440)-reflections around 525 °C and 400 °C, respectively. The 2θ Bragg peak positions of the characteristic (111)- and (440)-reflections were in good agreement with those obtained through conventional furnace annealing. Laser annealing of lithium manganese oxide films provided a quick and efficient technique and delivered a dominant (440)-reflection which showed the expected electrochemical behavior of the well-known two-step de-/intercalation process of lithium-ions into the cubic spinel structure within galvanostatic testing and cyclic voltammetry.

Aim: This article aims to evaluate the performance of a recent method to estimate heritability of continuous and binary traits, specifically in the context of pharmacogenetic studies. Materials & methods: The approach to be evaluated was designed to estimate heritability in large-scale disease studies. Extensive simulation studies designed to emulate common scenarios seen in pharmacogenetic studies were performed to elucidate the potential utility of this approach outside of disease genetics. The simulations cover continuous and binary traits with small-to-moderate heritability values across a variety of samples sizes in genome-wide, as well as candidate gene, settings. Results: On a genome-wide scale, a combination of relatively large sample sizes (i.e., n ≥ 1000) and at least moderate underlying heritability (i.e., ≥0.25) are needed in order to attain reasonable statistical power. However, in candidate gene studies, reasonable power can be attained across a more broad range of scenarios. Conclusion: Our simulation studies show that the proposed approach has clear utility in the context of pharmacogenetic studies, especially in candidate gene settings, and provides novel supplementary information that can be used to inform decision-making in the pharmaceutical industry. Original submitted 1 November 2012; Revision submitted 24 January 2013

Ultrasonographic (US) assessment of abdominal aortic aneurysms is typically performed by measuring maximal aneurysm diameter from two-dimensional images. These measurements are prone to inaccuracies owing to image planes and interobserver variability. The purpose of this study was to compare the variability in diameter, cross-sectional area (CSA), and volume measurements of abdominal aortic aneurysms obtained using a three-dimensional (3D) US imaging system with those obtained using computed tomographic (CT) angiography, and to determine the reliability of these measures.Seven patients in whom endovascular aneurysm repairs were performed underwent CT angiography in addition to a 3D US scan. Measurements computed using 3D surface reconstructions of CT and 3D US scans included maximum diameter, CSA, and aneurysm volume. The seven matched CT and 3D US scans were compared at baseline and 6 to 8 weeks later.The average aneurysm measured 57.2 mm on CT and 56.2 mm on US (P = 0.14). Correlation coefficients for diameter, CSA, and volume were 0.88, 0.90, and 0.93, respectively (all P values0.001). A Bland-Altman analysis demonstrated a strong agreement between 92% of the diameter, 96.4% of the CSA, and 100% of the volume measurements. The interrater reliability was remarkably high comparing the modalities (CT vs. US), and ranged from 0.934 to 0.997 for single measurements and 0.965 to 0.998 for all measurements together; moreover, there was a strong reliability when the tests were reviewed 6 to 8 weeks later, with a reliability of 0.962 to 0.998 for single measurements and 0.992 to 0.999 for all tests (all P values0.001).The 3D US is an accurate and noninvasive method of determining aneurysm size and geometry that is reproducible. Volumetric measurements may represent a significant advancement in long-term follow-up after endovascular aneurysm repair.

The WZ production cross section is measured by the CMS experiment at the CERN LHC in proton proton collision data samples corresponding to integrated luminosities of 4.9 fb(-1) collected at root s = 7 TeV, and 19.6fb(-1) at root s = 8 TeV. The measurements are performed using the fully-leptonic WZ decay modes with electrons and muons in the final state. The measured cross sections for 71 < m(Z) < 111 GeV are sigma (pp -> WZ; root s = 7 TeV) = 20.14 +/- 1.32 (stat)+/- 0.38 (theo)+/- 1.06 (exp)+/- 0.44 (lumi) pb and sigma (pp -> WZ; root s = 8 TeV) = 24.09 +/- 0.87 (stat) 0.80 (theo) +/- 1.40 (exp) +/- 0.63 (lumi) pb. Differential cross sections with respect to the Z boson p(T), the leading jet p(T), and the number of jets are obtained using the root s = 8 TeV data. The results are consistent with standard model predictions and constraints on anomalous triple gauge couplings are obtained.