Gut microbiota is associated with metabolic health in children with obesity.

BACKGROUND AND AIMS: We aimed to describe and characterize the gut microbiota composition and diversity in children with obesity according to their metabolic health status. METHODS: Anthropometry, Triglycerides, HDL cholesterol, HOMA-IR, and systolic and diastolic blood pressure (SBP, DBP) were evaluated (and z-score calculated) and faecal samples were collected from 191 children with obesity aged from 8 to 14. All children were classified depending on their cardiometabolic status in either a "metabolically healthy" (MHO; n = 106) or "metabolically unhealthy" (MUO; n = 85) group. Differences in gut microbiota taxonomies and diversity between groups (MUO vs MHO) were analysed. Alpha diversity index was calculated as Chao1 and Simpson's index, and ß-diversity was calculated as Adonis Bray-Curtis index. Spearman's correlations and logistic regressions were performed to study the association between cardiometabolic health and the microbiota. RESULTS: Children in the MUO presented significantly lower alpha diversity and richness than those in the MHO group (Chao1 index p = 0.021, Simpson's index p = 0.045, respectively), whereas microbiota ß-diversity did not differ by the cardiometabolic health status (Adonis Bray-Curtis, R2 = 0.006; p = 0.155). The MUO group was characterized by lower relative abundances of the genera Christensenellaceae R7 group (MHO:1.42% [0.21-2.94]; MUO:0.47% [0.02-1.60], p < 0.004), and Akkermansia (MHO:0.26% [0.01-2.19]; MUO:0.01% [0.00-0.36], p < 0.001) and higher relative abundances of Bacteroides (MHO:10.6% [4.64-18.5]; MUO:17.0% [7.18-27.4], p = 0.012) genus. After the adjustment by sex, age, and BMI, higher Akkermansia (OR: 0.86, CI: 0.75-0.97; p = 0.033), Christensenellaceae R7 group (OR: 0.86, 95% CI: 075-0.98; p = 0.031) and Chao1 index (OR: 0.86, CI: 0.96-1.00; p = 0.023) represented a lower risk of the presence of one or more altered cardiovascular risk factors. CONCLUSION: Lower proportions of Christensenellaceae and Akkermansia and lower diversity and richness seem to be indicators of a metabolic unhealthy status in children with obesity.

Exercise during pregnancy and its impact on mothers and offspring in humans and mice.

Exercise during pregnancy has beneficial effects on maternal and offspring's health in humans and mice. The underlying mechanisms remain unclear. This comparative study aimed to determine the long-term effects of an exercise program on metabolism, weight gain, body composition and changes in hormones [insulin, leptin, brain-derived neurotrophic factor (BDNF)]. Pregnant women (n=34) and mouse dams (n=44) were subjected to an exercise program compared with matched controls (period I). Follow-up in the offspring was performed over 6 months in humans, corresponding to postnatal day (P) 21 in mice (period II). Half of the mouse offspring was challenged with a high-fat diet (HFD) for 6 weeks between P70 and P112 (period III). In period I, exercise during pregnancy led to 6% lower fat content, 40% lower leptin levels and an increase of 50% BDNF levels in humans compared with controls, which was not observed in mice. After period II in humans and mice, offspring body weight did not differ from that of the controls. Further differences were observed in period III. Offspring of exercising mouse dams had significantly lower fat mass and leptin levels compared with controls. In addition, at P112, BDNF levels in offspring were significantly higher from exercising mothers while this effect was completely blunted by HFD feeding. In this study, we found comparable effects on maternal and offspring's weight gain in humans and mice but different effects in insulin, leptin and BDNF. The long-term potential protective effects of exercise on biomarkers should be examined in human studies.

Impaired DNase1-mediated degradation of neutrophil extracellular traps is associated with acute thrombotic microangiopathies.

BACKGROUND: Acute thrombotic microangiopathies (TMAs) are characterized by excessive microvascular thrombosis and are associated with markers of neutrophil extracellular traps (NETs) in plasma. NETs are composed of DNA fibers and promote thrombus formation through the activation of platelets and clotting factors. OBJECTIVE: The efficient removal of NETs may be required to prevent excessive thrombosis such as in TMAs. To test this hypothesis, we investigated whether TMAs are associated with a defect in the degradation of NETs. METHODS AND RESULTS: We show that NETs generated in vitro were efficiently degraded by plasma from healthy donors. However, NETs remained stable after exposure to plasma from TMA patients. The inability to degrade NETs was linked to a reduced DNase activity in TMA plasma. Plasma DNase1 was required for efficient NET degradation and TMA plasma showed decreased levels of this enzyme. Supplementation of TMA plasma with recombinant human DNase1 restored NET-degradation activity. CONCLUSIONS: Our data indicate that DNase1-mediated degradation of NETs is impaired in patients with TMAs. The role of plasma DNases in thrombosis is, as of yet, poorly understood. Reduced plasma DNase1 activity may cause the persistence of pro-thrombotic NETs and thus promote microvascular thrombosis in TMA patients.

Leptin does not induce an inflammatory response in the murine placenta.

Leptin is described as a pro-inflammatory signal in fat tissue, which is released from adipocytes and in turn activates immune cells. Also, leptin levels are known to be increased in pregnancies complicated with enhanced inflammatory processes in the placenta. Hence, we assumed that increased leptin amounts might contribute to inducing an inflammatory response in the placenta. To test this hypothesis, pregnant mice were continuously infused with recombinant murine leptin s. c. from day g13 to g16, resulting in a 3-fold increase of maternal circulating serum leptin levels. Dissected placentas were examined for the expression of pro-inflammatory cytokines IL-6 and TNF-alpha and the anti-inflammatory cytokine IL-10 using qPCR analysis. No changes were found except for TNF-alpha, which was slightly elevated upon leptin stimulation. However, TNF-alpha protein levels were not significantly higher in placentas from leptin treated mice. Also, leukocyte infiltration in the labyrinth section of placentas was not increased. In summary, our data demonstrate for the first time that elevated leptin levels alone do not induce an inflammatory response in the placenta.

Simultaneous online SPE-HPLC-MS/MS analysis of docetaxel, temsirolimus and sirolimus in whole blood and human plasma.

Docetaxel and temsirolimus are some of the most used drugs in a wide range of solid tumors. In preclinical studies, mTOR inhibitors such as temsirolimus have demonstrated synergistic cytotoxic effects with taxanes providing the rationale for combination studies. These anticancer agents exhibit a narrow therapeutic concentration range and due to their high inter- and intra-individual pharmacokinetic variability, therapeutic dose monitoring by highly sensitive methods as LC-MS/MS are important for clinical research. Therefore, the aim of this study was to develop and validate a sensitive, fast and convenient method for the simultaneous identification and quantification of docetaxel, temsirolimus and its main metabolite, sirolimus, using paclitaxel, another anticancer drug, as the internal standard. These analytes were quantified by an integrated online solid phase extraction-high performance liquid chromatography-tandem mass spectrometry (SPE-HPLC-MS/MS) system. Separation was performed on a Zorbax eclipse XDB-C8 (150mm×4.6mm, 5µm) column. The mass spectrometer tandem quadruple detector was equipped with jet stream electrospray ionization, monitored in multiple reactions monitoring (MRM) and operated in positive mode. A combination of protein precipitation with methanol/zinc sulphate (70:30) (v/v) and online SPE using a Zorbax eclipse plus C8 (12.5mm×4.6mm, 5µm) cartridge was used to extract the compounds. This method allows the use of the same reagents, sample treatment and analytical technique independently of whether the samples are whole blood or plasma. The method has been successfully validated and applied to real samples. It is a suitable method for dose adjustment and for evaluating potential drug interactions during combined treatments.

Changes in subthalamic activity during movement observation in Parkinson's disease: is the mirror system mirrored in the basal ganglia?

OBJECTIVE: The observation of a voluntary movement executed by another person is associated with an alpha and beta EEG desynchronization over the motor cortex, thought to reflect activity from the human "mirror neuron" system. The aim of our work was to study the changes in local field potentials (LFP) recorded from the subthalamic nucleus (STN) and their relationship with cortical activity, during movement observation. METHODS: Bilateral EEG and STN LFP recordings were acquired in 18 patients with Parkinson's disease, through surgically implanted electrodes for deep brain stimulation. Oscillatory changes during movement execution and movement observation were compared with two different control conditions (simple stimulus and rotating stimulus observation), in "off" and "on" motor states. Time-frequency transforms and event-related coherence were used for the analysis. RESULTS: Movement observation was accompanied by bilateral beta reduction in subthalamic power and cortico-STN coherence, which was smaller than the decrease observed during movement execution, but significant when compared with the two control conditions. CONCLUSIONS: Movement observation is accompanied by changes in the beta oscillatory activity of the STN, similar to those observed in the EEG. SIGNIFICANCE: These changes suggest that the basal ganglia might be engaged by the activity of the human mirror system.

[Brain oscillations: pathophysiological and potentially therapeutic role in some neurological and psychiatric diseases]. / Oscilaciones cerebrales: papel fisiopatológico y posiblemente terapéutico en algunas enfermedades neurológicas y psiquiátricas.

The terms "oscillations" or "oscillatory activity" are frequently used not only to define the rhythmic fluctuations of the postsynaptic potentials of a neuronal group (local field potentials) or a cortical region (EEG, MEG), but also to indicate the rhythmic discharge pattern of action potentials from a neuron or a small group of neurons. Oscillatory activity makes possible the synchronization of different neuronal groups from nearby or distant cortical regions that participate in the same motor, sensory or cognitive task. The presence of oscillatory activity is usually associated to the existence of synchronization, but both phenomena are not necessarily always equivalent. Abnormalities of oscillatory activities or synchronization within or between different brain structures have been described in several neurological and psychiatric diseases; these abnormalities might play a relevant pathophysiological role in Parkinson's disease (and other movement disorders), schizophrenia or epilepsy. This review discusses all these aspects, with emphasis on their potential role both as a basic mechanism in brain function and as a pathophysiological substrate for some of the symptoms and signs observed in several diseases.

Chirp-evoked potentials in the awake and anesthetized rat. A procedure to assess changes in cortical oscillatory activity.

Steady-state potentials are oscillatory responses generated by rhythmic stimulation of a sensory pathway. The frequency of the response, which follows the frequency of stimulation and potentially indicates the preferential working frequency of the auditory neural network, is maximal at a stimulus rate of 40 Hz for auditory stimuli in humans, but may be different in other species. Our aim was to explore the responses to different frequencies in the rat. The stimulus was a tone modulated in amplitude by a sinusoid with linearly-increasing frequency from 1 to 250 Hz ("chirp"). Time-frequency transforms were used for response analysis in 12 animals, awake and under ketamine/xylazine anesthesia. We studied whether the responses were due to increases in amplitude or to phase-locking phenomena, using single-sweep time-frequency transforms and inter-trial phase analysis. A progressive decrease in the amplitude of the response was observed from the maximal values (around 15 Hz) up to the limit of the test (250 Hz). The high-frequency component was mainly due to phase-locking phenomena with a smaller amplitude contribution. Under anesthesia, the amplitude and phase-locking of lower frequencies (under 100 Hz) decreased, while the phase-locking over 200 Hz increased. In conclusion, amplitude-modulation following responses differ between humans and rats in response range and frequency of maximal amplitude. Anesthesia with ketamine/xylazine modifies differentially the amplitude and the phase-locking of the responses. These findings should be taken into account when assessing the changes in cortical oscillatory activity related to different drugs, in healthy rodents and in animal models of neurodegenerative diseases.

Phenolic components and antioxidant activity of Fernblock, an aqueous extract of the aerial parts of the fern Polypodium leucotomos.

Fernblock, an aqueous extract of the aerial parts of the fern Polypodium leucotomos, used as raw material for topical and oral photoprotective formulations, was fractioned by HPLC and the main components with antioxidant capability were identified by means of UV spectra, electrochemical detection, and MSn. Phenolic compounds were identified as 3,4-dihydroxybenzoic acid, 4-hydroxybenzoic acid, vanillic acid, caffeic acid, 4-hydroxycinnamic acid, 4-hydroxycinnamoyl-quinic acid, ferulic acid, and five chlorogenic acid isomers. Total ferric antioxidant capacity (FRAP) of HPLC eluted fractions was measured. The results suggest that the herein identified compounds support, at least partially, the antioxidant and radical scavenging capacities of Fernblock.

[Abdominal aortic injury as a complication of laparoscopic cholecystectomy]. / Lesión de la aorta abdominal como complicación de la colecistectomía laparoscópica.

A 58-year-old woman with a history of high blood pressure and cesarean section by median laparotomy underwent elective laparoscopic cholecystectomy under general anesthesia. After initiation of pneumoperitoneum, hemodynamic instability developed (blood pressure 75/40 mm Hg, heart rate 47 beats/min) and resolved after volume replacement and administration of vasoactive drugs. The rest of the procedure was completed without further adverse events. After extubation, the patient was transferred to the anesthetic recovery ward, where she tended to hypotension that was unresponsive to volume replacement, with a hematocrit of 18% and hemoglobin concentration of 6.9 g/dL. Hemoperitoneum was suspected, and an emergency median laparotomy revealed 300 cc of blood in the peritoneal cavity, a large hematoma putting pressure on retroperitoneal structures, and 2 infrarenal aortic lesions (anterior and posterolateral). During the reoperation the patient required multiple transfusions and vasoactive drugs to maintain hemodynamic stability. Recovery in the postanesthetic care unit was uneventful. After transfer to the ward, on the 20th day after surgery she developed septic shock with intraabdominal fluid requiring surgical drainage and admission to the intensive care unit. Abdominal aortic injury during laparoscopic surgery usually occurs upon introduction of trocars for initiating pneumoperitoneum. The technique must be performed correctly and monitored properly to assure early detection of adverse events associated with high mortality.

[Hemothorax as complication of laparoscopic technique]. / Un hemotórax como complicación de la técnica laparoscópica.

A 69-year-old woman with a hiatal hernia and esophagitis caused by reflux was scheduled for laparoscopic Nissen fundoplication. Anesthetic induction was accomplished with fentanyl, propofol and rocuronium and maintenance with sevoflurane and rocoronium. Hemodynamics were stable until the end of surgery, when a sudden decrease in blood pressure to 40/20 mm Hg and arterial carbon dioxide tension to 14 mm Hg were observed. Anesthetic insufflation was stopped and physiological saline solution and ephedrine were administered. Hemothorax and air embolism were ruled out. After hemodynamic recovery, renewal of anesthetic insufflation was contraindicated, laparoscopy was halted, and hemoperitoneum was ruled out. After extubation, the patient was transferred to the recovery room, where hemodynamic instability worsened progressively. A chest radiograph suggest left hemothorax and an emergency thoracotomy was performed. After evacuation of multiple clots from the pleural cavity, the point of hemorrhage was located at the right crus of the diaphragm and bleeding was arrested. Postoperative recovery continued with no further events. The most common complications of laparoscopy are pneumothorax, gastroesophageal perforation and hemorrhage. However, given the complexity of the technique, the possibility of bleeding in the pleural cavity, as occurred in this case, should be considered. This case highlights the importance of rigorous intraoperative monitoring and postoperative surveillance by the anesthesiologist.

Photochemical reactivity of 1-substituted-1-aza-1,4-dienes promoted by electron-acceptor sensitizers. Di-pi-methane rearrangements and alternative reactions via radical-cation intermediates.

Irradiation of a series of beta,gamma-unsaturated imines, oxime acetates, and oxime methyl ethers, using 9,10-dicyanoanthrathene (DCA) or dicyanodurene (DCD) as electron acceptor sensitizers, affords the corresponding cyclopropanes resulting from 1-aza-di-pi-methane rearrangements via radical cations. In some cases, alternative reactions of these intermediates occur to yield nitriles, dihydroquinolines, dihydronaphthalene derivatives, and cycloaddition products. Some of these products result from reactions via alkene radical-cation intermediates while others arise by pathways involving imine radical-cation intermediates. The yields of products formed in these processes were significantly higher when DCD was used as electron-acceptor sensitizer instead of DCA.

[Use of acid-suppressive medications in hospitalized patients]. / Utilización de supresores de la secreción ácida en pacientes hospitalizados.

INTRODUCTION: The acid-suppressive medications are used frequently in hospitalized patients, but some studies suggests that their are overused, especially in the treatment of stress ulcer prophylaxis in low-risk patients. This research determine the frequency of use and indications of acid-suppressive medications in hospitalized patients in a internal medical service. MATERIALS AND METHODS: A at random and retrospective revision of 209 medical records of 661 hospitalized patients during the year 2000 in the V Internal Medical Service of the General University Hospital of Gregorio Marañón (Cantoblanco). DEFINITIONS: Use: any prescription of an acid-suppressive medication, regardless of dosage regimen, in which the patient received at least one dose. Appropriated indication: endoscopical criteria: treatment of active ulcer disease, esophagitis or gastritis. Medical criteria: symptomatic gastroesophageal reflux, dyspepsia, prolonging of previous treatment, as a part of a treatment regimen for H. pylori, relief of dyspepsia caused by nonsteroidal anti-inflammatory drugs, stress ulcer prophylaxis in high risk patients. Inappropriate indication: any other uses were considered not indicated. RESULTS: Of the 209 patients, a total of 157 (75.1%) received acid-suppressive therapy, average age 76 years old, 89 women (56.7%) and 68 men (43.3%). The admitting diagnoses for the admission were: CHF 34 (21.7%), Pneumonia (16.6%), UTI (15.3%) y COPD 18 (11.5%). The use in hospital ward was 84.7% (133), with inappropriate use 72.2% (96), the cause of prescription "no reason" 52.1% (50), and received proton pump inhibitors 46.9% (45). CONCLUSIONS: This study show a high frequency of incorrect use of acid-suppressive medications in hospitalized patients.

Simple high-performance liquid chromatography method for alpha-tocopherol measurement in Rosmarinus officinalis leaves. New data on alpha-tocopherol content.

A simple HPLC method for vitamin E (alpha-tocopherol) measurement in the leaves of Rosmarinus officinalis has been developed and validated. It has enabled new data for alpha-tocopherol content to be established. The leaves, recently harvested, were dried in a microwave oven and crushed; then, alpha-tocopherol was directly extracted from portions of ground material with acetone, by probe sonication. After centrifugation the acetonic extract was analysed by HPLC with ergocalciferol (vitamin D2) added as internal standard and a gradient elution with a Nucleosil C18 column at 35 degrees C. Validation parameters of the method can be considered adequate. For standards: linearity is r=0.999, recovery is 100+/-2%, intra-assay precision has RSD=+/-3% and inter-assay precision has RSD=+/-6%. For samples: linearity is r=0.99, recovery: 93+/-7%, intra-assay precision has RSD=+/-4% and inter-assay precision has RSD=+/-7%.