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.

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.

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.