In vitro effects of nonesterified fatty acids on bovine neutrophils oxidative burst and viability

An in vitro study was conducted to examine the influence of nonesterified fatty acids (NEFA) on bovine polymorphonuclear leukocytes (PMN). Eight healthy, midlactating Holstein cows were used as blood donors. Blood PMN were isolated and incubated with a mixture of NEFA, reflecting composition of bovine plasma NEFA at concentrations that were intended to mimic those found in blood of cows undergoing high, moderate, or low lipomobilization intensity (2, 1, 0.5, 0.25, 0.125, and 0.0625 mM). Control samples were incubated in absence of NEFA. Phagocytosis and oxidative burst activities were assessed by a 2-color flow cytometric method, which was based on oxidation of intracellular dihydrorhodamine 123 to green fluorescent rhodamine 123. Oxidative burst products were generated by incubating PMN with Staphylococcus aureus labeled with propidium iodide. A flow cytometric technique was used to detect PMN viability, necrosis, and apoptosis using fluorescein isothiocyanate-labeled annexin-V and propidium iodide. Phagocytic activity was not affected by NEFA. The highest concentration of NEFA (2 mM) was associated with a dramatic increase of phagocytosis-associated oxidative burst activities with a reduction in cell viability (48.0 vs. 97.5% in control samples) and with a marked increase of necrosis (49.4 vs. 0.5% in control samples). Conversely, the mixture of NEFA did not affect the occurrence of apoptosis. Enhancement of the oxidative burst associated with the highest concentration of NEFA might explain the reduced viability and higher percentage of necrosis observed under the same conditions. This study demonstrated a substantial resistance of bovine PMN to an overload of fatty acids. However, observation that the highest concentration of NEFA regulated some PMN functions encourages the possibility of in vivo studies to assess the relationships between intensity of lipomobilization, plasma NEFA, and bovine PMN functions

Urinary chitinase 3-like protein 1 for early diagnosis of acute kidney injury : a prospective cohort study in adult critically ill patients

Background: Acute kidney injury (AKI) occurs frequently and adversely affects patient and kidney outcomes, especially when its severity increases from stage 1 to stages 2 or 3. Early interventions may counteract such deterioration, but this requires early detection. Our aim was to evaluate whether the novel renal damage biomarker urinary chitinase 3-like protein 1 (UCHI3L1) can detect AKI stage >= 2 more early than serum creatinine and urine output, using the respective Kidney Disease vertical bar Improving Global Outcomes (KDIGO) criteria for definition and classification of AKI, and compare this to urinary neutrophil gelatinase-associated lipocalin (UNGAL). Methods: This was a translational single-center, prospective cohort study at the 22-bed surgical and 14-bed medical intensive care units (ICU) of Ghent University Hospital. We enrolled 181 severely ill adult patients who did not yet have AKI stage >= 2 based on the KDIGO criteria at time of enrollment. The concentration of creatinine (serum, urine) and CHI3L1 (serum, urine) was measured at least daily, and urine output hourly, in the period from enrollment till ICU discharge with a maximum of 7 ICU-days. The concentration of UNGAL was measured at enrollment. The primary endpoint was the development of AKI stage >= 2 within 12 h after enrollment. Results: After enrollment, 21 (12 %) patients developed AKI stage >= 2 within the next 7 days, with 6 (3 %) of them reaching this condition within the first 12 h. The enrollment concentration of UCHI3L1 predicted the occurrence of AKI stage >= 2 within the next 12 h with a good AUC-ROC of 0.792 (95 % CI: 0.726-0.849). This performance was similar to that of UNGAL (AUC-ROC of 0.748 (95 % CI: 0.678-0.810)). Also, the samples collected in the 24-h time frame preceding diagnosis of the 1st episode of AKI stage >= 2 had a 2.0 times higher (95 % CI: 1.3-3.1) estimated marginal mean of UCHI3L1 than controls. We further found that increasing UCHI3L1 concentrations were associated with increasing AKI severity. Conclusions: In this pilot study we found that UCHI3L1 was a good biomarker for prediction of AKI stage >= 2 in adult ICU patients

Development of a cytometric bead array screening tool for the simultaneous detection of pro-inflammatory cytokines in plasma of lipopolysaccharide-challenged pigs

Introduction : Lipopolysaccharide (LPS) has been widely used as a model of immune challenge in pigs as it induces the immediate synthesis of pro-inflammatory cytokines such as tumor necrosis factor-alpha (TNF-α), interleukin-1β (IL-1β) and IL-6. In research, multiplex assays currently are a very popular tool for the simultaneous detection of biomarkers of infection and inflammation. Specific and sensitive Enzyme-Linked Immuno Sorbent Assays (ELISAs) are well-suited to perform single factor analysis, yet for multi-parameter analyses, this approach is time-consuming and expensive. Cytometric bead array (CBA) is a flexible, bead-based flow cytometric application for the simultaneous detection of various soluble proteins of interest. The aim of the present study was to develop and validate a CBA 3-plex assay for the major pro-inflammatory cytokines TNF-α, IL-1β and IL-6. The results were compared to commercial ELISA kits. Materials and Methods : Four pigs with a mean body weight (BW) of 24.9 kg were intravenously challenged with 15 µg ultrapure LPS/kg BW (Escherichia coli serotype O111:B4). Plasma was isolated and stored at -70 °C until analysis. Capture antibodies were covalently coupled to the surface of beads with unique fluorescence intensities (Becton Dickinson Biosciences). Detection antibodies were conjugated with R-Phycoerythrin (R-PE). A mixture of beads was firstly incubated with an appropriate standard mixture. Subsequently, a mixture of detection antibodies, either directly or indirectly conjugated to R-PE, was added to accomplish the desired sandwich format. The samples were finally analyzed on a BD FACSArrayTM Bioanalyzer. ELISAs were purchased from R&D Systems. Results : Table 1 shows the validation parameters of the developed CBA 3-plex assay and the commercial ELISAs. Following an in vivo LPS challenge, similar plasma concentration-time profiles were observed for all cytokines with CBA and ELISA. Discussion : This is the first CBA study describing a validated multiplex protocol for the simultaneous measurement of the major porcine pro-inflammatory cytokines TNF-α, IL-1β and IL-6. In research, ELISAs are still considered as the gold standard for determination of secreted proteins in serum or plasma, however, the novel validated CBA 3-plex assay provides a fast and economical screening tool for determination of cytokine profiles in small porcine plasma volumes

Le cinéma érotique, panorama contemporain

Réalité et représentation, vérité et mensonge, tel est le quadrilatère où se sont longtemps décidé des limites de l\u27admissible en matière de sexe à l\u27écran. Mais ces limites sont précisément celles sur lesquelles ne cesse de jouer l\u27art au XXe s., notamment au cinéma où l\u27 « effet de réel » trouble les lignes. Où positionner la bibliothèque

Psychometric properties of the Chinese (Putonghua) version of the Oxford Cognitive Screen (OCS-P) in subacute poststroke patients without neglect

Background Oxford Cognitive Screen is designed for assessing cognitive functions of poststroke patients.This study was aimed to assess the psychometric properties of the Chinese (Putonghua) version of theOxford Cognitive Screen-Putonghua (OCS-P) for use among poststroke patientswithout neglect. Methods Expert reviewpanel evaluated content validity of theChinese-translated items. After pilot tested the translated items, the patients and healthy participants completed the OCS-P as well as theMontreal Cognitive Assessment (MoCA-ChiB) andGoldenberg’s test.Agroup of patients completedOCS-P for the second timewithin seven days.Data analyses included confirmatory factor analysis, item difficulty and item-total correlation, inter- and intrarater reliability, internal consistency, and between-group discrimination. Results One hundred patients and 120 younger ( = 60) or older ( = 60) healthy participants completed all the tests. Modifications were required for items in the “Picture Naming”, “Orientation”, and “Sentence Reading” subscales.Confirmatory factor analysis revealed a three-factor structure for theOCS-P subscales.The internal consistency coefficients for the three identified test dimensions were 0.30 to 0.52 (Cronbach’s alpha). Construct validity coefficients between the OCS-P and MoCA-ChiB subscales were 0.45 Conclusion This study generated satisfactory evidence on the content validity, substantive validity, construct validity, inter- and intrarater reliability, and known-group discrimination of theOCS-P.They support its application among poststroke patients who speak Putonghua. Future studies could review the existing five-dimension domains for improving its structural validity and internal consistency as well as generate evidence of the OCS-P for use among the poststroke patients with neglect.</p