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Introduction We evaluated the diagnostic reliability of serum polymerase chain reaction (PCR) versus blood culture, abdominal fluid or both (composite measure) in patients receiving empirical antifungal treatment for suspected invasive candidiasis. Methods This observational, prospective, non-interventional, multicentre study in Spain enrolled 176 critically ill patients admitted to the intensive care unit. Separate blood samples for culture and serum PCR were taken before the start of antifungal therapy. Patient assessment was performed according to each site’s usual clinical practice. The primary end point was concordance between serum PCR and blood culture. Secondary end points were concordance between serum PCR and a positive abdominal fluid sample or the composite measure. Quality indices included sensitivity, specificity, positive/negative predictive values (PPV/NPV) and kappa indices. Results Among 175 evaluable patients, rates of Candida detection were similar for serum PCR (n = 16/175, 9.1%) versus blood culture (n = 14/175, 8.0%). Quality indices for serum PCR relative to blood culture were: sensitivity 21.4%; specificity 91.9%; PPV 18.8%; NPV 93.1%; kappa index 0.125. Thirty-two abdominal fluid samples were positive. Quality indices for serum PCR versus abdominal fluid were: sensitivity 31.3%; specificity 83.0%; PPV 15.6%; NPV 92.3%; kappa index 0.100. Quality indices for serum PCR versus the composite measure were: sensitivity 15.8%; specificity 92.7%; PPV 37.5%; NPV 79.9%; kappa index 0.107. Conclusion The sensitivity of serum PCR for Candida detection was low and the rate of concordance was low between serum PCR and the other diagnostic techniques used to identify Candida infections. Hospital-based diagnostic tests need optimising to improve outcomes in patients with suspected invasive candidiasis. Funding Astellas Pharma Inc. Electronic supplementary material The online version of this article (10.1007/s40121-019-0248-z) contains supplementary material, which is available to authorized users.

Aims. To assess the possible effect of polyphenol-rich olive extracts on lipid metabolism in medaka fish by quantifying the expression of lipogenic and lipolytic genes. Materials and methods. Adult medaka fish were maintained in tanks for five days with five extracts at 0.01% in water, causing obesity through a diet rich in carbohydrates, with a control group maintained in water with a normal diet. The extracts contained polyphenols ranging between 7 and 116 mg/g (oleuropein, hydroxytyrosol) with an antioxidant power of 2&ndash, 13 mmol of 2,4,6-tri(2-pyridyl)-1,3,5-triazine/100 g. After five days, the fish were sacrificed and the hepatic mRNA and its complementary DNA were extracted by reverse transcription. Complementary DNAs were quantified for three lipolytic and three lipogenic genes by real-time PCR. The relative gene expression was calculated from the amplification curves in reference to the control group. Results. The expression of genes involved in lipolysis, including peroxisome proliferator-activated receptor-&plusmn, acyl-CoA oxidase 1, and carnitine palmitoyltransferase 1, were clearly decreased in fish subjected to an obesogenic diet, and this situation could not be reversed in fish maintained with polyphenol-rich extracts. In contrast, lipogenic fatty acid synthase, acetyl-CoA carboxylase 1, and sterol regulatory element-binding protein 1 genes increased considerably with the obesogenic diet and reverted to the normal state with the olive extracts. The effect was not dependent on the total polyphenol content, the specific oleuropein or hydroxytyrosol concentration, or the antioxidant power, suggesting a synergistic effect. Conclusion. Olive polyphenols, acting as anti-lipogenic agents, have a positive effect on lipid metabolism, but their mechanism in each gene is different according to the extract, which supports synergistic mechanisms with the different proportions of polyphenols and accompanying phytochemicals in each extract.

Objective: We evaluated the protective activity of an extract from a by-product such as olive stones, through its ability to inhibit H202 induced apoptosis in the SH-SY5Y human neuroblastoma cell line. Material and methods: To such end, 20,000 cells/well were cultivated and differentiation with retinoic acid was initiated. Once the cells were differentiated, apoptosis was induced with and without H2O2 extract. Finally, cDNA extraction was performed, and pro-apoptotic genes Bax and anti-apoptotic genes Bcl-2 were analyzed. Quantification of the gene expression was performed using the GAPDH gene marker. Results: Cell viability with the extract is 97.6% (SD 5.7) with 10 mg/l and 62.8% (SD 1.2) to 50 mg/l, using 10 mg/l for the biomarker assay. The retinoic acid differentiated SH-S cell line (10 μM) shows a clear apoptosis when treated with H2O2 150 μM, with a Bax/Bcl-2 ratio of 3.75 (SD 0.80) in contrast to the differentiated control cells subjected to H2O2 and with extract, which have the same ratio of 1.02 (SD 0.01-0.03). Conclusion: The olive stone extract shows anti-apoptotic activity in the provoked cell death of SH-SY5Y human neuroblastoma cells in their normal state, defending them from oxidative stress which produces a significant increase in the apoptotic gene ratio in contrast to anti-apoptotic genes (Bax/Bcl-2). Objetivo: evaluar la actividad protectora del extracto de un subproducto como son los huesos de aceitunas, mediante su capacidad de inhibir la apoptosis en la línea celular humana de neuroblastoma SH-SY5Y inducida con H2O2. Material y métodos: se han cultivado 20.000 cel/pocillo, iniciando diferenciación con ácido retinoico y, una vez diferenciadas las células, se ha inducido la apoptosis con H2O2 con extracto y sin presencia del mismo. Finalmente se efectúa la extracción de cDNA y el análisis de los genes proapoptótico Bax y antiapoptótico Bcl-2. La cuantificación de la expresión génica se realiza frente al gen marcador GAPDH. Resultados: la viabilidad celular con el extracto es del 97,6% (SD 5,7) con 10 mg/l y 62,8% (SD 1,2) a 50 mg/l, utilizando 10 mg/l para el ensayo de biomarcadores. Las células de la línea SH-S diferenciadas con ácido retinoico (10 μM), muestran una clara apoptosis al ser tratadas con H2O2 150 μM, con una relación Bax/Bcl2 de 3,75 (SD 0,80) frente a las células diferenciadas control y sometidas a H2O2 y con extracto que tienen la misma relación de 1,02 (SD 0,01-0,03). Conclusión: el extracto de huesos de aceitunas presenta una actividad antiapoptótica frente a la provocación de la muerte celular por peróxido de hidrógeno, preservando a las células de neuroblastoma humano SH-SY5Y en su estado de normalidad, al defenderlas del estrés oxidativo que produce un significativo aumento de la relación de genes apoptóticos frente a antiapoptóticos (Bax/Bcl2).

The administration of different polyphenols protects against increased body weight and fat accumulation. The aim of the study was to determine the anti-adipogenic activity of an olive-seed polyphenolic extract, by means of mouse fibroblast cell line 3T3-L1 adipocyte differentiation.Cells were incubated and differentiated (6000 cells/cup) in the presence of olive-seed extract at 10 and 50 mg/l biosecure concentrations of polyphenols, and with no extract in the control sample. After 5 to 7 days mature adipocytes are formed. The fat clusters are quantified by means of red-oil staining, 490 nm absorbance, and the expression of the leptin and PPARg genes, and then compared to the values obtained in the cultures before and after adipocyte differentiation.The control samples, with no extract, presented an accumulation of fat of 100%. By contrast, the addition of 50 mg/l of olive-seed extract polyphenols resulted in a 50% accumulation of fat, similar to that of the non-differentiated cells. A 10 mg/l extract concentration had no effect. Anti-adipogenic activity is thus confirmed, as the expression of the PPARg and leptin genes is reduced in adipocyte differentiation in the presence of extract at 50 mg/l. In conclusion, both the formation of fatty substances characteristic of adipogenesis, and the expression of the adipogenic PPARg and leptin genes are found to be inhibited by the prior addition of olive-seed extract polyphenols at a 50 mg/l concentration.La administración de diferentes polifenoles protege contra el incremento de peso y la acumulación de grasa. Objetivo: comprobar la actividad anti-adipogénica de un extracto polifenólico de huesos de aceituna, utilizando la diferenciación a adipocitos de la línea celular 3T3-L1 de fibroblastos de ratón. Material y métodos: se cultivan y diferencian las células (6.000 células/pocillo) en presencia del extracto de huesos de aceitunas a 10 y 50 mg/l de polifenoles, concentraciones bioseguras, y sin extracto como control. A los 5-7 días se forman los adipocitos maduros. Se cuantifican los cúmulos de grasa formados mediante tinción con Oil- Red y medida de la absorbancia a 490 nm y la expresión de los genes de leptina y PPARg, relacionándolos con los valores en los cultivos antes y después de diferenciarse a adipocitos. Resultados: las muestras control, sin extracto, se consideran el 100% de acumulación de grasas. En contraste, la adición de 50 mg/l de extracto de polifenoles de huesos de aceituna muestra un cúmulo de grasa de alrededor del 50%, semejante a las células no diferenciadas. Con 10 mg/l de extracto no se muestra efecto. Se confirma la actividad antiadipogénica, observándose disminución en la expresión de los genes PPARg y de leptina en la diferenciación a adipocitos en presencia del extracto a 50 mg/l. En conclusión, la formación de los cuerpos grasos característicos de la adipogénesis queda inhibida previa adición de 50 mg/l de polifenoles de extracto de huesos de aceituna, así como la expresión de los genes adipogénicos PPARg y de leptina.

We report a 48-year-old male admitted to hospital due to a severe alcoholic pancreatitis. At four weeks of evolution of the acute episode, an abdominal CAT scan showed a fluid collection of 20 cm diameter located in the pancreatic tail and 2 small collections in the head. The patient received several antimicrobials and during the seventh week of evolution, while receiving vancomycin, presented fever. A fine needle aspiration of the cyst revealed the presence of Haemophilus parainfluenzae biotype VIII. The patient was treated with amoxicillin-clavulanic acid and a laparoscopic cysto-gastrostomy, with a good clinical response.

Background The objective of this study is to analyze the factors that are associated with the adequacy of empirical antibiotic therapy and its impact in mortality in a large cohort of patients with extended-spectrum β-lactamase (ESBL) - producing Escherichia coli and Klebsiella spp. bacteremia. Methods Cases of ESBL producing Enterobacteriaceae (ESBL-E) bacteremia collected from 2003 through 2008 in 19 hospitals in Spain. Statistical analysis was performed using multivariate logistic regression. Results We analyzed 387 cases ESBL-E bloodstream infections. The main sources of bacteremia were urinary tract (55.3%), biliary tract (12.7%), intra-abdominal (8.8%) and unknown origin (9.6%). Among all the 387 episodes, E. coli was isolated from blood cultures in 343 and in 45.71% the ESBL-E was multidrug resistant. Empirical antibiotic treatment was adequate in 48.8% of the cases and the in hospital mortality was 20.9%. In a multivariate analysis adequacy was a risk factor for death [adjusted OR (95% CI): 0.39 (0.31-0.97); P = 0.04], but not in patients without severe sepsis or shock. The class of antibiotic used empirically was not associated with prognosis in adequately treated patients. Conclusion ESBL-E bacteremia has a relatively high mortality that is partly related with a low adequacy of empirical antibiotic treatment. In selected subgroups the relevance of the adequacy of empirical therapy is limited.

We report a 48-year-old male admitted to hospital due to a severe alcoholic pancreatitis. At four weeks of evolution of the acute episode, an abdominal CAT scan showed a fluid collection of 20 cm diameter located in the pancreatic tail and 2 small collections in the head. The patient received several antimicrobials and during the seventh week of evolution, while receiving vancomycin, presented fever. A fine needle aspiration of the cyst revealed the presence of Haemophilus parainfluenzae biotype VIII. The patient was treated with amoxicillin-clavulanic acid and a laparoscopic cysto-gastrostomy, with a good clinical response.

The objective of this study was to compare the activity of ciprofloxacin, levofloxacin, moxifloxacin, gatifloxacin and ofloxacin against Pseudomonas aeruginosa by determining the mutant prevention concentration (MPC).Thirty-four clinical isolates of Pseudomonas aeruginosa causing nosocomial infection were studied. MPC values were determined using an inoculum of 10(10) cfu/mL on Mueller-Hinton plates with serial dilutions of the antibiotics. The microorganisms were classified according to whether the patients had been previously treated with fluoroquinolones or not.The fluoroquinolone with the lowest MPC values was ciprofloxacin, followed by levofloxacin. Analysis of strains from patients previously treated with fluoroquinolones showed persistent susceptibility according to the CLSI criteria, but with lower activity of all the drugs.Although the microorganism remained susceptible to fluoroquinolones according to classical methods, the suitability of using these drugs in patients with severe infection and a history of fluoroquinolone use should be evaluated.