A novel Monoclonal Antibody against Notch1 Targets Leukemia-associated Mutant Notch1 and Depletes Therapy Resistant Cancer Stem Cells in Solid Tumors

Higher Notch signaling is known to be associated with hematological and solid cancers. We developed a potential immunotherapeutic monoclonal antibody (MAb) specific for the Negative Regulatory Region of Notch1 (NRR). The MAb604.107 exhibited higher affinity for the ``Gain-offunction'' mutants of Notch1 NRR associated with T Acute lymphoblastic Leukemia (T-ALL). Modeling of the mutant NRR with 12 amino-acid insertion demonstrated ``opening'' resulting in exposure of the S2-cleavage site leading to activated Notch1 signaling. The MAb, at low concentrations (1-2 mu g/ml), inhibited elevated ligand-independent Notch1 signaling of NRR mutants, augmented effect of Thapsigargin, an inhibitor of mutant Notch1, but had no effect on the wild-type Notch1. The antibody decreased proliferation of the primary T-ALL cells and depleted leukemia initiating CD34/CD44 high population. At relatively high concentrations, (10-20 mu g/ml), the MAb affected Notch1 signaling in the breast and colon cancer cell lines. The Notch-high cells sorted from solid-tumor cell lines exhibited characteristics of cancer stem cells, which were inhibited by the MAb. The antibody also increased the sensitivity to Doxorubucinirubicin. Further, the MAb impeded the growth of xenografts from breast and colon cancer cells potentiated regression of the tumors along with Doxorubucin. Thus, this antibody is potential immunotherapeutic tool for different cancers.

Structure of an amidohydrolase, SACOL0085, from methicillin-resistant Staphylococcus aureus COL

Staphylococcus aureus is an opportunistic pathogen that rapidly acquires resistance to frontline antibiotics. The characterization of novel protein targets from this bacterium is thus an important step towards future therapeutic strategies. Here, the crystal structure of an amidohydrolase, SACOL0085, from S. aureus COL is described. SACOL0085 is a member of the M20D family of peptidases. Unlike other M20D peptidases, which are either monomers or dimers, SACOL0085 adopts a butterfly-shaped homotetrameric arrangement with extensive intersubunit interactions. Each subunit of SACOL0085 contains two Mn2+ ions at the active site. A conserved cysteine residue at the active site distinguishes M20D peptidases from other M20 family members. This cysteine, Cys103, serves as bidentate ligand coordinating both Mn2+ ions in SACOL0085.

Draft genome of a commonly misdiagnosed multidrug resistant pathogen Candida auris

Background: Candida auris is a multidrug resistant, emerging agent of fungemia in humans. Its actual global distribution remains obscure as the current commercial methods of clinical diagnosis misidentify it as C. haemulonii. Here we report the first draft genome of C. auris to explore the genomic basis of virulence and unique differences that could be employed for differential diagnosis. Results: More than 99.5 % of the C. auris genomic reads did not align to the current whole (or draft) genome sequences of Candida albicans, Candida lusitaniae, Candida glabrata and Saccharomyces cerevisiae; thereby indicating its divergence from the active Candida clade. The genome spans around 12.49 Mb with 8527 predicted genes. Functional annotation revealed that among the sequenced Candida species, it is closest to the hemiascomycete species Clavispora lusitaniae. Comparison with the well-studied species Candida albicans showed that it shares significant virulence attributes with other pathogenic Candida species such as oligopeptide transporters, mannosyl transfersases, secreted proteases and genes involved in biofilm formation. We also identified a plethora of transporters belonging to the ABC and major facilitator superfamily along with known MDR transcription factors which explained its high tolerance to antifungal drugs. Conclusions: Our study emphasizes an urgent need for accurate fungal screening methods such as PCR and electrophoretic karyotyping to ensure proper management of fungemia. Our work highlights the potential genetic mechanisms involved in virulence and pathogenicity of an important emerging human pathogen namely C. auris. Owing to its diversity at the genomic scale; we expect the genome sequence to be a useful resource to map species specific differences that will help develop accurate diagnostic markers and better drug targets.

Corrosion behaviour of creep-resistant AE42 magnesium alloy-based hybrid composites developed for powertrain applications

The corrosion behaviour of AE42 magnesium alloy and its composites reinforced with Saffil short fibres and SiC particles in various combinations was investigated. The corrosion rate of the unreinforced alloy was the lowest. The composite reinforced with Saffil short fibre alone exhibited slightly lower corrosion rate than the hybrid composites containing both Saffil short fibres and SiC particles. However, there was no specific trend observed in the corrosion rate of the hybrid composites with respect to the SiC particle content. The degradation of corrosion resistance of the composites was mainly attributed to the irregular and loose surface films.

Evaluation of bioactive sphingolipids in 4-HPR-resistant leukemia cells

Background -- N-(4-hydroxyphenyl)retinamide (4-HPR, fenretinide) is a synthetic retinoid with potent pro-apoptotic activity against several types of cancer, but little is known regarding mechanisms leading to chemoresistance. Ceramide and, more recently, other sphingolipid species (e.g., dihydroceramide and dihydrosphingosine) have been implicated in 4-HPR-mediated tumor cell death. Because sphingolipid metabolism has been reported to be altered in drug-resistant tumor cells, we studied the implication of sphingolipids in acquired resistance to 4-HPR based on an acute lymphoblastic leukemia model. Methods -- CCRF-CEM cell lines resistant to 4-HPR were obtained by gradual selection. Endogenous sphingolipid profiles and in situ enzymatic activities were determined by LC/MS, and resistance to 4-HPR or to alternative treatments was measured using the XTT viability assay and annexin V-FITC/propidium iodide labeling. Results -- No major crossresistance was observed against other antitumoral compounds (i.e. paclitaxel, cisplatin, doxorubicin hydrochloride) or agents (i.e. ultra violet C, hydrogen peroxide) also described as sphingolipid modulators. CCRF-CEM cell lines resistant to 4-HPR exhibited a distinctive endogenous sphingolipid profile that correlated with inhibition of dihydroceramide desaturase. Cells maintained acquired resistance to 4-HPR after the removal of 4-HPR though the sphingolipid profile returned to control levels. On the other hand, combined treatment with sphingosine kinase inhibitors (unnatural (dihydro)sphingosines ((dh)Sph)) and glucosylceramide synthase inhibitor (PPMP) in the presence or absence of 4-HPR increased cellular (dh)Sph (but not ceramide) levels and were highly toxic for both parental and resistant cells. Conclusions -- In the leukemia model, acquired resistance to 4-HPR is selective and persists in the absence of sphingolipid profile alteration. Therapeutically, the data demonstrate that alternative sphingolipid-modulating antitumoral strategies are suitable for both 4-HPR-resistant and sensitive leukemia cells. Thus, whereas sphingolipids may not be critical for maintaining resistance to 4-HPR, manipulation of cytotoxic sphingolipids should be considered a viable approach for overcoming resistance.

Evaluation of wheat or corn dried distillers' grains with solubles on performance and carcass characteristics of feedlot steers

[EN]A study was conducted on crossbred steers (n=275; 376±924 kg) to evaluate performance and carcass quality of cattle fed wheat or corn dried distillers’ grains with solubles (DDGS). The control ration contained 86.6% rolled barley grain, 5.7% supplement and 7.7% barley silage (DM basis). The four treatments included replacement of barley grain at 20 or 40% of the diet (DM basis) with wheat or corn DDGS. Steers were slaughtered at a common end weight of 645 kg with 100 steers randomly (n=20 per treatment) selected for determination of the retail yield of sub-primal boneless boxed beef (SPBBB). Data were analyzed as a completely randomized design using pen as the experimental unit. Feeding increasing levels of wheat DDGS led to a quadratic increase in dry matter intake (DMI) (P<0.01), whereas increasing levels of corn DDGS led to a quadratic decrease in DMI (P=0.01). Average daily gain was not influenced (P=0.13) by feeding wheat or corn DDGS, but cattle fed corn DDGS exhibited a quadratic increase (P=0.01) in gain:feed. As a result, a quadratic increase (P<0.01) in calculated NEg of the diet was observed as corn DDGS levels increased. A linear decrease (P=0.04) in days on feed (169, 166 and 154 d) was noted when increasing levels of wheat DDGS (0, 20 and 40%) were fed. Dressing percentage increased in a linear fashion with wheat DDGS (P<0.01) inclusion level and in a quadratic fashion (P=0.01) as corn DDGS inclusion level increased although other carcass traits were not affected (P=0.10) by treatment. The results indicate that replacement of barley grain with corn or wheat DDGS up to 40% of the diet (DM) can lead to superior performance (improved gain:feed or reduced days on feed, respectively) with no detrimental effect on quality grade or carcass SPBBB yield.

The Costs of Drought: the Exceptional 2007-2008 Case of Barcelona

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Diseño y desarrollo de una solución de resistant communications al protocolo Host Identity Protocol

[ES]Este Trabajo de Fin de Grado consiste en diseñar y desarrollar una solución de resilient communications para su uso en entornos de movilidad, en concreto, en entornos vehiculares. Se diseñara una solución que consiste en añadir soporte de múltiples vías de comunicación entre dos extremos para el protocolo de movilidad HIP. Este trabajo consiste en buscar una solución de resilient communications, ya que buscamos como objetivo principal aumentar la disponibilidad del sistema de comunicaciones, es decir, aumentar aspectos tales como la tolerancia a fallos y contra ataques de seguridad, concretamente contra ataques contra la disponibilidad.

A humidity-resistant highly sensitive holographic photopolymerizable dry film

A new humidity-resistant highly sensitive acrylamide-based photopolymeric holographic recording material has been developed. The photopolymer is resistant to the humidity of environment. Diffraction efficiencies near 50% are obtained with exposure energy of 60 mJ/cm(2) in materials of 150 mu m. thickness. Diphenyl iodonium chloride is added to the material and can increase the exposure sensitivity by a factor of more than 4 (to about 28 mJ/cm(2)). An image has been successfully stored in the material with a small distortion. (C) 2005 Elsevier B.V. All rights reserved.

Carbapenem-resistant and carbapenem-susceptible isogenic isolates of Klebsiella pneumoniae ST101 causing infection in a tertiary hospital

Background: In this study we describe the clinical and molecular characteristics of an outbreak due to carbapenem-resistant Klebsiella pneumoniae (CR-KP) producing CTX-M-15 and OXA-48 carbapenemase. Isogenic strains, carbapenem-susceptible K. pneumoniae (CS-KP) producing CTX-M-15, were also involved in the outbreak. Results: From October 2010 to December 2012 a total of 62 CR-KP and 23 CS-KP were isolated from clinical samples of 42 patients (22 had resistant isolates, 14 had susceptible isolates, and 6 had both CR and CS isolates). All patients had underlying diseases and 17 of them (14 patients with CR-KP and 3 with CS-KP) had received carbapenems previously. The range of carbapenem MICs for total isolates were: imipenem: 2 to >32 mu g/ml vs. <2 mu g/ml; meropenem: 4 to >32 mu g/ml vs. <2 mu g/ml; and ertapenem: 8 to >32 mu g/ml vs. <2 mu g/ml. All the isolates were also resistant to gentamicin, ciprofloxacin, and cotrimoxazole. Both types of isolates shared a common PFGE pattern associated with the multilocus sequence type 101 (ST101). The bla(CTX-M-15) gene was detected in all the isolates, whereas the bla(OXA-48) gene was only detected in CR-KP isolates on a 70 kb plasmid. Conclusions: The clonal spread of K. pneumoniae ST101 expressing the OXA-48 and CTX-M-15 beta-lactamases was the cause of an outbreak of CR-KP infections. CTX-M-15-producing isolates lacking the blaOXA-48 gene coexisted during the outbreak.

River Ure: fisheries drought report 1996

Yorkshire Water Services (YWS) are currently granted a Time Limited Licence (TLL) for abstraction at Kilgram Bridge which is due for renewal in 1999. The Environment. Agency requires information on fish populations with regards to drought conditions and any possible effects that abstraction may have when considering licence renewal. In' order to evaluate any effects of drought and abstraction a three year study was instigated to examine fish populations. Surveys were conducted at nine main River Ure sites and two tributaries in which the triennial rolling programme formed the basis of site selection. Multi-method sampling techniques were carried out at several sites in order to evaluate capture efficiency. High densities of brown trout juveniles were observed in the tributaries with an indication that fish had become crowded as a result of low flows. Recruitment of brown trout in the tributaries was not directly related to flow levels in the main R. Ure. However, it is concluded that salmonids are at risk during drought flows and high temperatures from increased susceptibility to disease, predation, poor water quality and the direct lethal effect of high temperatures in shallow water.

River Ouse: fisheries drought report 1996

The basis for undertaking this study was to examine factors and potential impacts affecting fish and fishing in relation to low flow drought conditions and what other impacts may arise as a result of further reduced flows resulting from abstraction. The study formed the basis of a three year project to concentrate on effects relating to potable water abstractions at Moor Monkton by YWS. To fully evaluate the possible effects on fisheries the study set out to encompass fish population surveys from fry to adult stock, analysis of angler catch data, reports from anglers and river reports from Environment Agency Fisheries staff. In order to evaluate any effects of drought and abstraction a three year study was instigated to examine fish populations. Fish population surveys were conducted at six sites in which the triennial rolling programme formed the basis of site selection. Multi-method sampling techniques were carried out at several sites in order to evaluate capture efficiency. Roach were prolific above the weir at Linton-on-Ouse, with gudgeon, perch and small bream also well represented. Roach dominated catches on the R.Ouse below Linton, with perch and bleak also relatively abundant. Low flows were not thought to be directly correlated to successful recruitment of coarse fish, rather the associated high temperatures during drought conditions showing a strong positive effect with most species exhibiting growth rates above their long-term average. At this stage in the study there are no clear indications that the drought has caused any deleterious effects to coarse fish populations or marked changes in species composition, with evidence of good recruitment by several species, indicating that the higher temperatures have generally been beneficial to recruitment. However, the indication that dace did not benefit as well as other coarse fish under these conditions may suggest some species are affected more than others. The successful strong recruitment of most coarse fish suggests that, in future, fisheries will be supported by the 1995 year-class.

Environmental effects of drought and abstractions of River Wharfe fisheries, summer 1997

This investigation was carried out to provide information on fish stocks and angling activity during 1997 in relation to the drought and, in particular, flows as influenced by Time Limited Licences. These abstractions will be for review in 1999. This report extends and updates the data presented for 1996. Fish population surveys (including eels) were undertaken on the main river and selected tributaries. Angler caught brown trout were examined, angler catch data have been reviewed, and observations by Environment Agency fisheries staff collated. It appeared that in River Wharfe both the fish populations and individual fish appeared to be in good condition and limited changes had occurred since the 1996 survey.