Electroencephalogram-based indices applied to dogs' depth of anaesthesia monitoring.

Hypnotic drug administration causes alterations in the electroencephalogram (EEG) in a dose-dependent manner. These changes cannot be identified easily in the raw EEG, therefore EEG based indices were adopted for assessing depth of anaesthesia (DoA). This study examines several indices for estimating dogs' DoA. Data (EEG, clinical end-points) were collected from 8 dogs anaesthetized with propofol. EEG was initially collected without propofol. Then, 100 ml h⁻¹ (1000 mg h⁻¹) of propofol 1% infusion rate was administered until a deep anaesthetic stage was reached. The infusion rate was temporarily increased to 200 ml h⁻¹ (2000 mg h⁻¹) to achieve 80% of burst suppression. The index performance was accessed by correlation coefficient with the propofol concentrations, and prediction probability with the anaesthetic clinical end-points. The temporal entropy and the averaged instantaneous frequency were the best indices because they exhibit: (a) strong correlations with propofol concentrations, (b) high probabilities of predicting anaesthesia clinical end-points.

Treatment planning systems dosimetry auditing project in Portugal.

BACKGROUND AND PURPOSE: The Medical Physics Division of the Portuguese Physics Society (DFM_SPF) in collaboration with the IAEA, carried out a national auditing project in radiotherapy, between September 2011 and April 2012. The objective of this audit was to ensure the optimal usage of treatment planning systems. The national results are presented in this paper. MATERIAL AND METHODS: The audit methodology simulated all steps of external beam radiotherapy workflow, from image acquisition to treatment planning and dose delivery. A thorax CIRS phantom lend by IAEA was used in 8 planning test-cases for photon beams corresponding to 15 measuring points (33 point dose results, including individual fields in multi-field test cases and 5 sum results) in different phantom materials covering a set of typical clinical delivery techniques in 3D Conformal Radiotherapy. RESULTS: All 24 radiotherapy centers in Portugal have participated. 50 photon beams with energies 4-18 MV have been audited using 25 linear accelerators and 32 calculation algorithms. In general a very good consistency was observed for the same type of algorithm in all centres and for each beam quality. CONCLUSIONS: The overall results confirmed that the national status of TPS calculations and dose delivery for 3D conformal radiotherapy is generally acceptable with no major causes for concern. This project contributed to the strengthening of the cooperation between the centres and professionals, paving the way to further national collaborations.

Pontin is a critical regulator for AML1-ETO-induced leukemia.

The oncogenic fusion protein AML1-ETO, also known as RUNX1-RUNX1T1 is generated by the t(8;21)(q22;q22) translocation, one of the most frequent chromosomal rearrangements in acute myeloid leukemia (AML). Identifying the genes that cooperate with or are required for the oncogenic activity of this chimeric transcription factor remains a major challenge. Our previous studies showed that Drosophila provides a genuine model to study how AML1-ETO promotes leukemia. Here, using an in vivo RNA interference screen for suppressors of AML1-ETO activity, we identified pontin/RUVBL1 as a gene required for AML1-ETO-induced lethality and blood cell proliferation in Drosophila. We further show that PONTIN inhibition strongly impaired the growth of human t(8;21)(+) or AML1-ETO-expressing leukemic blood cells. Interestingly, AML1-ETO promoted the transcription of PONTIN. Moreover, transcriptome analysis in Kasumi-1 cells revealed a strong correlation between PONTIN and AML1-ETO gene signatures and demonstrated that PONTIN chiefly regulated the expression of genes implicated in cell cycle progression. Concordantly, PONTIN depletion inhibited leukemic self-renewal and caused cell cycle arrest. All together our data suggest that the upregulation of PONTIN by AML1-ETO participate in the oncogenic growth of t(8;21) cells.

Fuzzy logic model to describe anesthetic effect and muscular influence on EEG Cerebral State Index.

The well-known Cerebral State Index (CSI) quantifies depth of anesthesia and is traditionally modeled with Hill equation and propofol effect-site concentration (Ce). This work brings out two novelties: introduction of electromyogram (EMG) and use of fuzzy logic models with ANFIS optimized parameters. The data were collected from dogs (n=27) during routine surgery considering two propofol administration protocols: constant infusion (G1, n=14) and bolus (G2, n=13). The median modeling error of the fuzzy logic model with Ce and EMG was lower or similar than that of the Hill with Ce (p=0.012-G1, p=0.522-G2). Furthermore, there was no significant performance impact due to model structure alteration (p=0.288-G1, p=0.330-G2) and EMG introduction increased or maintained the performance (p=0.036-G1, p=0.798-G2). Therefore, the new model can achieve higher performance than Hill model, mostly due to EMG information and not due to changes in the model structure. In conclusion, the fuzzy models adequately describe CSI data with advantages over traditional Hill models.

Associating ECG features with firefighter's activities.

In this paper we associate features obtained from ECG signals with the expected levels of stress of real firefighters in action when facing specific events such as fires or car accidents. Five firefighters were monitored using wearable technology collecting ECG signals. Heart rate and heart rate variability features were analyzed in consecutive 5-min intervals during several types of events. A questionnaire was used to rank these types of events according to stress and fatigue and a measure of association was applied to compare this ranking to the ECG features. Results indicate associations between this ranking and both heart rate and heart rate variability features extracted in the time domain. Finally, an example of differences in inter personal responses to stressful events is shown and discussed, motivating future challenges within this research field.

Correlation between clinical signs of depth of anaesthesia and cerebral state index responses in dogs during induction of anaesthesia with propofol.

The cerebral state index (CSI) is used for monitoring EEG and depth of anaesthesia. The objective of this study was to analyse the correlation between ocular reflexes, CSI and estimated propofol plasma concentrations (PropCP) in dogs during induction of anaesthesia with propofol. Fourteen dogs were premedicated with acepromazine 0.05 mg kg(-1) IM. Anaesthesia was induced with a 200 ml h(-1) propofol 1% constant infusion rate until loss of corneal reflex using RugLoop II software with Beths' pharmacokinetic model to estimate PropCp. Palpebral reflex (PR) and the corneal reflex (CR) were tested every 30s and classified as present (+) or absent (-), and eyeball position was registered as rotated ventromedialy (ERV) or centred (EC). Heart rate (HR), mean arterial pressure (MAP) and CSI values were analyzed from baseline before the beginning of propofol infusion (T0) until loss of CR; CSI and PropCp, CSI and anaesthetic planes, and PropCp and anaesthetic planes were compared using correlation analysis. PropCp reached 7.65+/-2.1 microg ml(-1) at the end of the study. CSI values at T0 were 89.2+/-3.8. Based on the observation of ocular reflexes and eyeball position, it was possible to define five anaesthetic planes: A (superficial) to E (deep), being A (PR+/CR+/EC), B (PR+/ERV/CR+), C (PR-/ERV/CR+), D (PR-/EC/CR+) and E (PR-/EC/CR-). There was a significant correlation between PropCp and the anaesthetic planes (R=0,861; P<0.01). No significant correlation was observed between CSI and the anaesthetic planes or between CSI and PropCp. MAP decreased significantly from T0 until loss of corneal reflex (from 98+/-14 mm Hg to 82+/-12 mm Hg); HR did not change significantly (from 101+/-30 bpm to 113+/-16 bpm). The CSI monitoring was not consistent with the clinical observations observed in the different stages of depth anaesthesia. This could limit the use of CSI for monitoring depth of anaesthesia with propofol.

A step towards effect-site target-controlled infusion with propofol in dogs: a k(e0) for propofol.

Target-controlled infusion (TCI) anesthesia using target effect-site concentration rather than plasma concentration provides less drug consumption, safer anesthesia, less undesired side effects and improved animal welfare. The aim of this study was to calculate the constant that converts propofol plasma into effect-site concentration (k(e0)) in dogs, and to implement it in a TCI system and compare it with the effect on the central nervous system (CNS). All dogs were subjected to general anesthesia using propofol. Fourteen dogs were used as the pilot group to calculate k(e0), using the t(peak) method. Fourteen dogs were used as the test group to test and validate the model. RUGLOOP II software was used to drive the propofol syringe pump and to collect data from S/5 Datex monitor and cerebral state monitor. The calculated k(e0) was incorporated in an existing pharmacokinetic model (Beths Model). The relationship between propofol effect site concentrations and anesthetic planes, and propofol plasma and effect-site concentrations was compared using Pearson's correlation analysis. Average t(peak) was 3.1 min resulting in a k(e0) of 0.7230 min(-1). The test group showed a positive correlation between anesthetic planes and propofol effect-site concentration (R = 0.69; P < 0.0001). This study proposes a k(e0) for propofol with results that demonstrated a good adequacy for the pharmacokinetic model and the measured effect. The use of this k(e0) will allow an easier propofol titration according to the anesthetic depth, which may lead to a reduction in propofol consumption and less undesired side effects usually associated to high propofol concentrations in dogs.

The immunological synapse and Rho GTPases.

Rho GTPases are molecular switches controlling a broad range of cellular processes including lymphocyte activation. Not surprisingly, Rho GTPases are now recognized as pivotal regulators of antigen-specific T cell activation by APCs and immunological synapse formation. This review summarizes recent advances in our understanding of how Rho GTPase-dependent pathways control T lymphocyte motility, polarization and activation.

A 2-Mb YAC/BAC-based physical map of the ovum mutant (Om) locus region on mouse chromosome 11.

The embryonic lethal phenotype observed when DDK females are crossed with males from other strains results from a deleterious interaction between the egg cytoplasm and the paternal pronucleus soon after fertilization. We have previously mapped the Om locus responsible for this phenotype, called the DDK syndrome, to an approximately 2-cM region of chromosome 11. Here, we report the generation of a physical map of 28 yeast and bacterial artificial chromosome clones encompassing the entire genetic interval containing the Om locus. This contig, spanning approximately 2 Mb, was used to map precisely genes and genetic markers of the region. We determined the maximum physical interval for Om to be 1400 kb. In addition, 11 members of the Scya gene family were found to be organized into two clusters at the borders of the Om region. Two other genes (Rad51l3 and Schlafen 2) and one EST (D11Wsu78e) were also mapped in the Om region. This integrated map provides support for the identification of additional candidate genes for the DDK syndrome.

BALB/c alleles at modifier loci increase the severity of the maternal effect of the "DDK syndrome".

The Om locus was first described in the DDK inbred mouse strain: DDK mice carry a mutation at Om resulting in a parental effect lethality of F(1) embryos. When DDK females are mated with males of other (non-DDK) inbred strains, e.g., BALB/c, they exhibit a low fertility, whereas the reciprocal cross, non-DDK females x DDK males, is fertile (as is the DDK intrastrain cross). The low fertility is due to the death of (DDK x non-DDK)F(1) embryos at the late-morula to blastocyst stage, which is referred to as the "DDK syndrome." The death of these F(1) embryos is caused by an incompatibility between a DDK maternal factor and the non-DDK paternal pronucleus. Previous genetic studies showed that F(1) mice have an intermediate phenotype compared to parental strains: crosses between F(1) females and non-DDK males are semisterile, as are crosses between DDK females and F(1) males. In the present studies, we have examined the properties of mice heterozygous for BALB/c and DDK Om alleles on an essentially BALB/c genetic background. Surprisingly, we found that the females are quasi-sterile when mated with BALB/c males and, thus, present a phenotype similar to DDK females. These results indicate that BALB/c alleles at modifier loci increase the severity of the DDK syndrome.

Fibroblast growth factor 2 promotes pancreatic epithelial cell proliferation via functional fibroblast growth factor receptors during embryonic life.

Several investigators have postulated that soluble growth factors are involved in the early development of the pancreas. In many tissues in which soluble factors are implicated in development, these factors act on their target cells through tyrosine kinase receptors. Because we had some preliminary evidence that fibroblast growth factor receptors (FGFRs) were expressed in the early pancreas, we investigated the effect of fibroblast growth factors (FGFs) during embryonic pancreatic development. For that purpose, we first studied the distribution and the functionality of FGFRs during pancreatic organogenesis. FGFR1 and FGFR4 were shown to be expressed at a high level during early pancreatic development before embryonic day 16, their levels of expression decreasing thereafter. The functionality of FGFR was studied next. It was demonstrated in vitro that both FGF1 and FGF2 induce the expression of NGFI-A mRNA, a useful indicator of functional growth factor-signaling pathways. Finally, the effect of FGF2 on embryonic pancreatic epithelial cell proliferation was studied. It was shown that FGF2 induces the proliferation of pancreatic epithelial cells during embryonic life. Taken together, these data strongly suggest that FGFs are implicated in pancreatic development during embryonic life.

Effects of modified atmosphere on crop productivity and mineral content.

Wheat, potato, pea and tomato crops were cultivated from seeding to harvest in a controlled and confined growth chamber at elevated CO2 concentration (3700 microL L-1) to examine the effects on biomass production and edible part yields. Different responses to high CO2 were recorded, ranging from a decline in productivity for wheat, to slight stimulation for potatoes, moderate increase for tomatoes, and very large enhancement for pea. Mineral content in wheat and pea seeds was not greatly modified by the elevated CO2. Short-term experiments (17 d) were conducted on potato at high (3700 microL L-1) and very high (20,000 microL L-1) CO2 concentration and/or low O2 partial pressure (approximately 20,600 microL L-1 or 2 kPa). Low O2 was more effective than high CO2 in total biomass accumulation, but development was affected: Low O2 inhibited tuberization, while high CO2 significantly increased production of tubers.

Outdoor experimental ponds (mesocosms) designed for long-term ecotoxicological studies in aquatic environment.

Outdoor artificial ponds (mesocosms) of 12 m3 were designed for long-term ecotoxicological studies. Sediment, macrophytes (Typha angustifolia and Elodea canadensis), and free and caged freshwater snails [Lymnaea palustris (Müller)] and wood lice (Asellus aquaticus L.) were collected in nearby natural ecosystems and introduced in the mesocosms. Sixty goldfish (Carassius auratus L.) were caged in each pond. Introduced species developed and reproduced in every mesocosm. Animal species (mainly insects and amphibians) spontaneously colonized the ponds, developed, and reproduced. The resulting communities qualitatively resemble those living in natural lentic systems in the surrounding area. Homogenity in physical and chemical conditions and in abundance of phytoplanktonic, periphytic, and macroinvertebrate communities between the different mesocosms was assessed during the stabilization period (8 months). Except for periphyton biomass, no divergent evolution was observed between the ponds. Mesocosm water was slightly eutrophic, alkaline (mean pH: 8.47 +/- 0.09), and moderately hard and mineralized. The homogenous and realistic environmental conditions and high ecological representativity of the outdoor experimental ponds were suitable for extensive ecotoxicological studies. Considerations on the choice and origin of introduced species and on possible interactive effects of the transfer of organisms from natural environments, maintainance conditions, and pollutant exposure are discussed.

Neurotrophin-3 increases intracellular calcium in a rat insulin-secreting cell line through its action on a functional TrkC receptor.

Pancreatic beta cells and neuronal cells show a large number of similarities. For example, functional receptors for nerve growth factor are present in beta cells. Here we investigate whether TrkC, a neuronal high affinity receptor for neurotrophin-3, is expressed in the insulin-secreting cell line INS-1. We demonstrate the expression in INS-1 cells of mRNAs coding for TrkC identical in size to those found in the brain. As in neuronal cells, different alternatively spliced forms of TrkC mRNA, differing by the insertion of an alternative exon in their kinase domain, were expressed in INS-1 cells. TrkC protein is also expressed in INS-1 cells and is functional. Indeed, when INS-1 cells were treated with neurotrophin-3, TrkC became phosphorylated on tyrosine residues, and the expression of early response genes was induced. This activation of the receptor was paralleled by a rapid and transient increase in cytosolic free calcium due to an influx of extracellular calcium. Functional receptors for NT-3 are thus expressed in INS-1 cells. This cell line provides a new model for the study of NT-3 signal transduction and should be useful in the understanding of the role of neurotrophins in insulin-secreting cells.