Transuterine Fetal Tracheal Occlusion Model in Mice

Fetal tracheal occlusion (TO), an established treatment modality, promotes fetal lung growth and survival in severe congenital diaphragmatic hernia (CDH). Following TO, retention of the secreted epithelial fluid increases luminal pressure and induces lung growth. Various animal models have been defined to understand the pathophysiology of CDH and TO. All have their own advantages and disadvantages such as the difficulty of the technique, the size of the animal, cost, high mortality rates, and the availability of genetic tools. Herein, a novel transuterine model of murine fetal TO is described. Pregnant mice were anesthetized, and the uterus exposed via a midline laparotomy. The trachea of selected fetuses were ligated with a single transuterine suture placed behind the trachea, one carotid artery, and one jugular vein. The dam was closed and allowed to recover. Fetuses were collected just before parturition. Lung to body weight ratio in TO fetuses was higher than that in control fetuses. This model provides researchers with a new tool to study the impact of both TO and increased luminal pressure on lung development

SURAT PENCATATAN CIPTAAN Karya Rekaman: Inovasi Pengajaran Gerak Dasar Tari Bali Dengan Bahasa Inggris Dalam Upaya Memperkokoh Kiprah ISI Denpasar di Dunia Internasional

Spatial joins are join operations that involve spatial data types and operators. Spatial access methods are often used to speed up the computation of spatial joins. This paper addresses the issue of benchmarking spatial join operations. For this purpose, we first present a WWW-based benchmark generator to produce sets of rectangles. Using a Web browser, experimenters can specify the number of rectangles in a sample, as well as the statistical distributions of their sizes, shapes, and locations. Second, using the generator and a well-defined set of statistical models we define several tests to compare the performance of three spatial join algorithms: nested loop, scan-and-index, and synchronized tree traversal. We also added a real-life data set from the Sequoia 2000 storage benchmark. Our results show that the relative performance of the different techniques mainly depends on two parameters: sample size, and selectivity of the join predicate. All of the statistical models and algorithms are available on the Web, which allows for easy verification and modification of our experiments.Peer Reviewe

Cell necrosis, intrinsic apoptosis and senescence contribute to the progression of exencephaly to anencephaly in a mice model of congenital chranioschisis

Amniotic fluid; Neonatal mortality; ExencephalyLíquido amniótico; Mortalidad neonatal; ExencefaliaLíquid amniòtic; Mortalitat neonatal; ExencefàliaExencephaly/anencephaly is one of the leading causes of neonatal mortality and the most extreme open neural tube defect with no current treatments and limited mechanistic understanding. We hypothesized that exencephaly leads to a local neurodegenerative process in the brain exposed to the amniotic fluid as well as diffuse degeneration in other encephalic areas and the spinal cord. To evaluate the consequences of in utero neural tissue exposure, brain and spinal cord samples from E17 exencephalic murine fetuses (maternal intraperitoneal administration of valproic acid at E8) were analyzed and compared to controls and saline-injected shams (n = 11/group). Expression of apoptosis and senescence genes (p53, p21, p16, Rbl2, Casp3, Casp9) was determined by qRT-PCR and protein expression analyzed by western blot. Apoptosis was measured by TUNEL assay and PI/AV flow cytometry. Valproic acid at E8 induced exencephaly in 22% of fetuses. At E17 the fetuses exhibited the characteristic absence of cranial bones. The brain structures from exencephalic fetuses demonstrated a loss of layers in cortical regions and a complete loss of structural organization in the olfactory bulb, hippocampus, dental gyrus and septal cortex. E17 fetuses had reduced expression of NeuN, GFAP and Oligodendrocytes in the brain with primed microglia. Intrinsic apoptotic activation (p53, Caspase9 and 3) was upregulated and active Caspase3 localized to the layer of brain exposed to the amniotic fluid. Senescence via p21-Rbl2 was increased in the brain and in the spinal cord at the lamina I-II of the somatosensory dorsal horn. The current study characterizes CNS alterations in murine exencephaly and demonstrates that degeneration due to intrinsic apoptosis and senescence occurs in the directly exposed brain but also remotely in the spinal cord.This work was supported by Prof. Jose L. Peiro internal Cincinnati Children's Hospital funding

Fetal Tracheal Occlusion Increases Lung Basal Cells via Increased Yap Signaling

Basal cell; Fetal tracheal occlusion; MechanotransductionCélula basal; Oclusión traqueal fetal; MecanotransducciónCèl·lula basal; Oclusió traqueal fetal; MecanotransduccióFetal endoscopic tracheal occlusion (FETO) is an emerging surgical therapy for congenital diaphragmatic hernia (CDH). Ovine and rabbit data suggested altered lung epithelial cell populations after tracheal occlusion (TO) with transcriptomic signatures implicating basal cells. To test this hypothesis, we deconvolved mRNA sequencing (mRNA-seq) data and used quantitative image analysis in fetal rabbit lung TO, which had increased basal cells and reduced ciliated cells after TO. In a fetal mouse TO model, flow cytometry showed increased basal cells, and immunohistochemistry demonstrated basal cell extension to subpleural airways. Nuclear Yap, a known regulator of basal cell fate, was increased in TO lung, and Yap ablation on the lung epithelium abrogated TO-mediated basal cell expansion. mRNA-seq of TO lung showed increased activity of downstream Yap genes. Human lung specimens with congenital and fetal tracheal occlusion had clusters of subpleural basal cells that were not present in the control. TO increases lung epithelial cell nuclear Yap, leading to basal cell expansion.Funding was obtained from NIH/NHLBI R01HL141229 (to BV)

Cell necrosis, intrinsic apoptosis and senescence contribute to the progression of exencephaly to anencephaly in a mice model of congenital chranioschisis

Exencephaly/anencephaly is one of the leading causes of neonatal mortality and the most extreme open neural tube defect with no current treatments and limited mechanistic understanding. We hypothesized that exencephaly leads to a local neurodegenerative process in the brain exposed to the amniotic fluid as well as diffuse degeneration in other encephalic areas and the spinal cord. To evaluate the consequences of in utero neural tissue exposure, brain and spinal cord samples from E17 exencephalic murine fetuses (maternal intraperitoneal administration of valproic acid at E8) were analyzed and compared to controls and saline-injected shams (n = 11/group). Expression of apoptosis and senescence genes (p53, p21, p16, Rbl2, Casp3, Casp9) was determined by qRT-PCR and protein expression analyzed by western blot. Apoptosis was measured by TUNEL assay and PI/AV flow cytometry. Valproic acid at E8 induced exencephaly in 22% of fetuses. At E17 the fetuses exhibited the characteristic absence of cranial bones. The brain structures from exencephalic fetuses demonstrated a loss of layers in cortical regions and a complete loss of structural organization in the olfactory bulb, hippocampus, dental gyrus and septal cortex. E17 fetuses had reduced expression of NeuN, GFAP and Oligodendrocytes in the brain with primed microglia. Intrinsic apoptotic activation (p53, Caspase9 and 3) was upregulated and active Caspase3 localized to the layer of brain exposed to the amniotic fluid. Senescence via p21-Rbl2 was increased in the brain and in the spinal cord at the lamina I-II of the somatosensory dorsal horn. The current study characterizes CNS alterations in murine exencephaly and demonstrates that degeneration due to intrinsic apoptosis and senescence occurs in the directly exposed brain but also remotely in the spinal cord

Biases in precipitation records found in parallel measurements

Presentación realizada en: 10th EUMETNET Data Management Workshop celebrado en St. Gallen, Suiza, del 28 al 30 de octubre de 2015.In this work we investigate biases introduced by the transition from Conventional to automatic precipitation measurements. This is another study in the framework of The Parallel Observations Scientific Team (POST, http://www.surfacetemperatures.org/databank/parallel_measurements), which is a newly created group of the International Surface Temperature Initiative (ISTI) supported by the World Meteorological Organization (WMO). The goals of POST are the study of climate data inhomogeneities at the daily and sub-daily level. Long instrumental climate records are usually affected by non-climatic changes, due to various reasons like relocations, changes in instrumentation, measurements schemes etc. Such inhomogeneities may distort the climate signal and can influence the assessment of trends and variability. For studying climatic changes it is important to accurately distinguish non-climatic from climatic signals. This can be achieved by studying the differences between two parallel measurements. These need to be sufficiently close together to be well correlated. One important ongoing worldwide transition is the one from manual to automated measurements. We need to study the impact of automated measurements urgently because sooner or later this will affect most of the stations in individual national networks. Similar to temperature series, we study the transition from conventional manual measurements (CON) to Automatic Weather Stations (AWS), using several parallel datasets distributed over Europe and America. The ratio series AWS-CON are subject to quality control, and before the analysis obvious errors are removed. Further, the series are inspected for internal inhomogeneities and– if necessary –the records are split into two or more homogeneous segments. Finally, each segment is studied to understand the biases introduced by the transition, its seasonality as well as changes in the empirical distributions. When additional variables are available, an attempt is made to study the effects of other variables on the observed biases

Description of the bias introduced by the transition from Conventional Manual Measurements to Automatic Weather Station through the analysis of European and American parallel datasets (+ Australia, Israel & Kyrgyzstan)

Presentación realizada en: 10th EUMETNET Data Management Workshop celebrado en St. Gallen, Suiza, del 28 al 30 de octubre de 2015.In this work, we approach the description of the biases introduced by automation in temperature records. This is one of the first studies in the framework of The Parallel Observations Scientific Team (POST). POST is a newly created group of the International Surface Temperature Initiative (ISTI), with the support of the World Meteorological Organization (WMO). The goals of POST (http://www.surfacetemperatures.org/databank/parallel_measurements) are the study of climate data inhomogeneities at the daily and sub-daily level through the compilation and analysis of parallel measurements. Long instrumental climate records are usually affected by non-climatic changes, due to, e.g., relocations and changes in instrumentation, instrument height or data collection and manipulation procedures. These so-called inhomogeneities distort the climate signal and can hamper the assessment of trends and variability. Thus to study climatic changes we need to accurately distinguish non-climatic and climatic signals. The most direct way to study the influence of non-climatic changes on the distribution and to understand the reasons for these biases is the analysis of parallel measurements. A parallel measurement is composed of two or more time series, which measure a climatic variable with two different systems (for example, Montsouris and Stevenson Screens) or in two different locations (for example, city centre and airport). They mimic the situation “before” and “after” a homogeneity break. Most parallel measurements are obtained from collocated or nearly collocated series and can help us to understand the size and shape of different typical sources of inhomogeneity, which affect the climate series. Here we study the transition from conventional temperature manual measurements (CON) to Automatic Weather Stations (AWS), using several parallel datasets distributed over Europe and America. The variables studied in the analysis presented here are daily maximum and minimum temperature. First of all, the metadata – when available - is gathered to gain knowledge on the exact setting of the parallel series. Secondly, the difference (temperature) series AWS-CON are submitted to quality control, to remove obvious errors and inspected to detect internal inhomogeneities and split if necessary. In a third step, each segment is studied to understand the bias introduced by the transition, its seasonality as well as changes in the empirical distributions. When additional variables are available, an attempt is made to study the effects of other variables on the observed biases.With the support of Grant CGL2012-32193, Ministerio de Economía y Competitividad, MINECO, España and FP7-SPACE-2013-1 grand 607193, Uncertainties in Ensembles of Regional Reanalyses (UERRA)

Alteracions funcionals de la via motora central en l’encefalopatia hepàtica experimental

En pacients cirròtics s’ha demostrat alteracions funcionals de la via cortico-espinal amb estimulació magnètica transcortical, consistents en una disminució de l’amplitud dels potencials evocats motors i en un increment de la latència. Aquestes alteracions funcionals s’associen a un increment de la senyal T2 per ressonància magnètica, probablement per un increment d’aigua cerebral. Les alteracions motores funcionals reverteixen amb el transplant hepàtic. a) En el primer treball s’ha demostrar alteracions funcionals en la via motora en dos models animals d’insuficiència hepàtica: cirrosi per tetraclorur de carboni i anastomosi porto-cava Les troballes reprodueixen les observades en humans, pel que podria ser útil en el desenvolupament d’un model experimental . No obstant, van ser trobades sota la influència d’anestèssics. Donat que la insuficiència hepàtica pot induir canvis en el metabolisme dels anestèssics, vem tenir interès per desenvolupar un model que no utilitzi anestèssics. b) En el segon estudi es va desenvolupar una nova metodologia per realitzar els estudis neurofisiològics en animals desperts, sense cap tipus d’anestèssia, i en lliure moviment. La nova metodologia està basada en la implantació de 6 elèctrodes subcutanis per poder fer els registres. Es va demostrar que la implantació dels eslèctrodes es poden mantenir com a mínim 3 setmanes per estudis repetits. A més a més, per validar el mètode, demostrem l’efecte dosi-depenent de tres diferents anestèssics sobre els potencials evocats motors. Aquesta nova metodologia pot fer servei per l’estudi en altres malaties com lesions medulars sense que l’anestèssia jugui un paper important. c) En el tercer estudi es va aplicar la nova metodologia sense anestèssia a diferents models de lesió hepàtica. Es va demostrar que les alteracions en els models de derivació porto-sistèmica i cirrosi es devien als efectes dels anestèssics. Per altra banda s’ha demostrat que el mètode és vàlid per mesurar l’encefalopatia hepàtica i de forma objectiva en models d’encefalopatia hepàtica aguda per insuficiènica hepàtica i encefalopatia hepàtica induïda per sang en el tub digestiu en rates amb anastomosis porto-cava. Aquesta nova metodologia té un gran valor científic, ja que l’encefalopatia hepàtica no es podia mesurar de cap manera objectiva, ja que tots els estudis anteriors es basaven en alteracions delscicle circadians, alteracions de comportament o aprenentatge que es mesuren amb mètodes no objectius. En canvi amb la neurofisiologia, és pot fer una mesura objectiva sense cap efecte per part de l’investigador i d’una manera quantificable. d) El quart estudi ha mostrat que és possible veure l’efecte d’un fàrmac que disminueix l’amoníac plasmàtic en rates amb anastomosis porto-cava que reben sang al tub digestiu. Aquest fàrmac, anomenat ornitina-fenilacetat (OP), ha començat a ser estudiat en pacients amb cirrosi hepàtica. La present tesis a resultat molt útil per obrir una nova línia: els estudis de nous fàrmacs per l’encefalopatia hepàtica en un model experimental. La tesi inclou el desenvolupament d’una nova metodologia, la validació d’un model i l’estudi dels mecanismes d’acció d’un fàrmac per l’encefalopatia hepàtica. Els resultats tenen una aplicabilitat immediata: -el model ha estat incorporat a la cartera de serveis de l’Institut de Recerca Hospital Vall Hebron. -El fàrmac estudiat es farà servir en pacients amb encefalopatia hepàtica.Cirrhotic patients exhibit functional abnormalities in the corticospinal tract by transcartical magnetic stimulation, these patients show a decrease on the amplitude of the motor evoked potentials and an increase on the latency. These functional alterations are associated with an increase in the T2 signal by magnetic resonance, probably by an increase of the water content. The motor abnormalities return to normal after hepatic transplant. a) In the first study of the thesis, we demonstrate functional abnormalities of the motor tract in two animal models of hepatic disease: cirrhosis induced by carbon tetrachloride and portacaval anastomosis. These findings reproduce the human abnormalities and can be useful for the development of an animal model. However, this study was performed under anhestesia, since the hepatic failure can induce changes in the anesthetics metabolism; we are interested in the development of a model without anesthesia. b) In the second study we developed a new methodology to perform neurophysiological studies in awake animals, without any anesthesia, and in free movement. The new methodology is based on subcutaneous implantation of 6 electrodes to record the motor evoked potentials. It was demonstrated that the introduction of electrodes can keep at least 3 weeks for repeated studies. In addition, to validate the method, we show dose-dependent effect of three different anesthetics on motor evoked potentials. This new methodology can service the study in other diseases such as cord injury without the anesthesia play an important role. c) In the third study we applied the new methodology without anesthesia in different models of hepatic injury. It was demonstrated that alterations in portacaval anastomosis and cirrhosis were due to the effects of anesthetics. On the other hand has shown that the method is valid for measuring hepatic encephalopathy objectively in models of acute hepatic encephalopathy by acute liver failure and hepatic encephalopathy induced by blood in the digestive tract in rats with portacaval anastomosis. This new methodology has a scientific value, as hepatic encephalopathy could not be measured in any objective way, since all previous studies were based on changes in the circadian rhythms; changes in behavior or learning are measured with no objectives methods. In contrast with the neurophysiology, motor evoked potentials is an objective and a quantifiable way not affected by the investigator. d) The fourth study showed that it is possible to see the effect of a drug that lowers plasma ammonia in rats with portocaval anastomosis receiving blood in the digestive tract. This drug, called ornithine-phenylacetate (OP), has begun to be studied in patients with liver cirrhosis. The present results are very useful for opening a new line: studies of new drugs for hepatic encephalopathy in an experimental model. The thesis includes the development of a new methodology, the validation of a model study of the mechanisms of action of a drug for hepatic encephalopathy. The results have an immediate applicability: - The model has been incorporated into the service of Valle Hebron Hospital Research Institute. - The study drug will be used in patients with hepatic encephalopathy