Alzheimer's Disease and Related Dementias Volume 10 - 2018 | https://doi.org/10.3389/fnagi.2018.00218 This article is part of the Research TopicThe Role of Glia in Alzheimer’s DiseaseView all 11 articles Neuroimaging has become an unparalleled tool to understand the central nervous system (CNS) anatomy While an altered immune function and microglia hyperactivation are common neuropathological features for many CNS disorders and neurodegenerative diseases direct assessment of the role of microglial cells remains a challenging task including magnetic resonance imaging (MRI) positron emission tomography (PET) and single positron emission computed tomography (SPECT) are widely used for human clinical applications and a variety of ligands are available to detect neuroinflammation and minimally invasive multiphoton microcopy (MPM) provides high resolution detection of single microglia cells we review in vivo real-time MPM approaches to assess microglia in preclinical studies including individual cell responses in surveillance protection and restoration of brain tissue integrity as well as in different pathological situations We focus on in vivo studies that assess the role of microglia in mouse models of Alzheimer’s disease (AD) analyzing microglial motility and recruitment as well as the role of microglia in anti-amyloid-β treatment MPM provides a high contrast and high spatial resolution approach to follow microglia chronically in vivo in complex models supporting MPM as a powerful tool for deep intravital tissue imaging An ideal neuroimaging technique would provide spatial resolution to allow subcellular morphological and physiological studies as well as high temporal resolution and sensitivity Labeled ligands and tracers used should also be characterized to the extent that the metabolism of the compound does not interfere with the sensitivity or specificity of the ligand while being able to cross the blood-brain barrier the half-life of radioactive compounds must be long enough to allow their quantification and provide a high signal-to-noise ratio Preclinical SPECT resolution may also reach <1 mm and about 8–12 mm in the case of clinical SPECT (Khalil et al., 2011) molecules labeled with positron emitting radionucleotides the resulting emission of detected γ-rays are used to image and measure biochemical processes in vivo the short half-lives of [15O] or [11C] require the on-site presence of a cyclotron to produce the radioisotopes Techniques employing tomographic reconstruction are used to generate three-dimensional images and it is possible to measure metabolic processes and explore brain pathophysiology as well as drug treatment responses The administration of exogenous contrast media allows the measurement of biological processes in some occasions it has been the development of multiphoton microscopy (MPM) that has significantly improved the possibility of deep (upto 1 mm) chronic in vivo imaging at the subcellular level Optical imaging precludes the need for radioactive ligands used in PET and SPECT and the large number of fluorescent ligands allows extremely diverse structural and functional readouts The main disadvantages are that the approach is invasive and the fact that only a limited portion of the brain can be assessed this is a very powerful approach for animal studies MPM has been largely used as a reference technique to explore the central nervous system (CNS) morphology and function in preclinical studies that include neural network activity immune system responses and the role of microglia progressive pathology or cellular responses in different pathological situations MPM is based on the probability that two or more low energy photons interact nearly simultaneously with a fluorescent molecule. This induces an electronic transition comparable to the absorption of one photon with double the energy. Then, a single photon is emitted by the excited fluorophore (Denk et al., 1990) By reaching <1 μm spatial resolution MPM allows cellular and subcellular discrimination without suffering from the slow image acquisition of MRI and PET Representative images of cranial window implantation and imaging setup for real time in vivo brain imaging with minimally invasive multiphoton microcopy (MPM) Cranial window surgeries have been performed with slightly different approaches and detailed protocols have been previously described (Mostany and Portera-Cailliau, 2008) the stereotaxic frame and all surgical surfaces are disinfected drill and glass coverslips are sterilized in a micro bead sterilizer Small pieces (~1.5 mm2) of gel foam are soaked in sterile saline to be used during the surgical procedure Animals are anesthetized with isoflurane: ~3% for induction and ~1% for the surgery while continuously monitoring animal reflexes Body temperature is maintained with a water recirculating blanket during the entire surgery-imaging procedure the sedated animal is placed in a stereotaxic frame and eye ointment is applied to prevent dry eyes The hair is trimmed in between the eyes from the neck to the eyes and the area is swapped with cotton tips dipped in iodopovidone and ethanol alternatively Local anesthesia (lidocaine) is subcutaneously injected and anti-inflammatory drugs (corticoids or non-steroid anti-inflammatory drugs) can also be administered before commencing the surgery With a scalpel and scissors the skin is opened and the muscles and periosteum are carefully removed to guarantee access to the cranium Initial drilling is performed to mark the cranium area between Bregma and Lambda to be removed (~6 mm in diameter) The skull is thinned in a circular pattern by careful drilling until it is almost detached At this moment the area is removed with fine forceps and wet gel foam is gently applied to the brain surface to limit swelling and bleeding fine forceps can be used to pull it gently from the craniotomy borders and leave it on the midline without damaging the leptomeningeal vessels The coverslip (8 mm in diameter) is dipped in sterile saline placed on top of the craniotomy and attached to the cranium with dental cement the animal receives subcutaneous opioids (buprenorphine) and acetaminophen is administered in the drinking water for the next three consecutive days or allowed recover from the surgery for several days or weeks A well-performed surgery leads to windows that allow longitudinal imaging for up to 1 year or even longer periods giving them a much more complex role in brain homeostasis and function Multiphoton in vivo tools for microglia imaging due to the fact that many more cells proliferate in the close proximity of a dying cell Real time in vivo multiphoton imaging of microglia in the cerebral cortex from CX3CR1-GFP mice Representative images of sequential follow up of a control mouse and two mice after parenchyma laser ablation for 10 and 30 s Microglia processes start going towards the lesion site as soon as 1 min after laser ablation and the processes continue extending towards the lesion 150 min later Vessels (blue) are filled with Texas Red dextran 70 KD green) and white arrows point to laser ablation sites Scale bar 50 μm and insets 12 μm All of these promising tools will help to evaluate microglial immunomodulation in physiological and pathological brain conditions the positive role of microglial activation in removing Aβ can be compromised by the concomitant effect of an increased secretion of proinflammatory compounds that might be toxic for nearby neurons Additional efforts are required to define the role of TREM2 in health and disease In depth analysis of microglia dynamics showed that the number of microglia increases over a month independent of the volume of senile plaques While larger plaques were surrounded by larger microglia the average microglia size appeared to be stable over time Real time in vivo multiphoton imaging of microglia in the cerebral cortex from APP/PS1×CX3CR1-GFP mice Representative images of microglia clusters around senile plaques (A,D) senile plaques are labeled with methoxy-XO4 (red) The scale bar is 50 μm and for the insets 10 μm Recently, Füger et al. (2017) have also followed the natural history of microglia in an AD mouse model (CD11b-CreERT2;R26-tdTomato;APPPS1 mice) These authors reported a higher microglia cell loss in AD mice (~20%) when compared with wildtype mice (~13%) at 10 months of age They also showed that cell division of non-plaque-associated microglia was over three times more common than microglia loss in APP/PS1 animals newly formed microglia were reported to move toward nearby amyloid plaques Since the rates of plaque-associated microglia disappearance and proliferation were similar the authors postulated that the increase in the numbers of microglia surrounding plaques results from the proliferation of microglia in plaque-free areas it cannot be ruled out that plaque-associated tdTomato-positive cells are derived it seems that microglial motility and recruitment with results depending on specific amyloid depositing transgenic mouse models the effect seems to be limited to older PDAPP mice (14–17 months of age) while no differences were observed in younger mice (3.5 months old) suggesting that the microglial response is only detected when aggregated Aβ is present it seems that senile plaques are a triggering factor to form microglia clusters and support the idea that while microglia do not seem to successfully clear plaques by themselves they might be activated by anti-Aβ antibodies and contribute to Aβ clearance the underlying mechanisms remain to be completely elucidated the high spatial resolution and limited penetration of the MPM technique precludes whole brain imaging significant training and expertise to successfully perform the surgeries and the ability to image and reimage the brains of mice requires a significant investment making it hard to detect whether the neuroinflammatory progress occurs early on or later during disease which is a primary aim of the study of microglia in vivo in vivo multiphoton imaging is a powerful approach to assess the role of microglia in AD It allows structural and functional imaging of the living brain Future studies exploiting this technique should be able to clearly delineate the normal and pathophysiological role of neuroinflammation in the brain increasing our understanding of the cellular and molecular changes during progression of disease All studies were conducted with approved protocols from the Massachusetts General Hospital Animal Care and Use Committee and in compliance with NIH guidelines for the use of experimental animals or approved by the Animal Care and Use Committee of the University of Cadiz in accordance with the Guidelines for Care and Use of experimental animals (European Commission Directive 2010/63/UE and Spanish Royal Decree53/2013) CH-B drafted part of the manuscript and reviewed it All authors contributed to and have approved the final manuscript Cultura y Deporte en el marco del Programa Estatal de Promoción del Talento y su Empleabilidad en I+D+Ii del Plan Estatal de Investigación Científica y Técnica y de Innovación 2013–2016 Salvador de Madariaga (PRX16/00246) Programa Estatal de I+D+I orientada a los Retos de la Sociedad (BFU 2016—75038-R) financed by the Agencia Estatal de Investigación (AEI) and the Fondo Europeo de Desarrollo Regional (FEDER) Ciencia y Empleo Junta de Andalucía (P11-CTS-7847) Subvención para la financiación de la investigación y la innovación biomédica y en ciencias de la salud en el marco de la iniciativa territorial integrada 2014–2020 para la provincia de Cádiz financed by the Fondo de Desarrollo Regional (FEDER; PI-0008-2017) The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest Evaluation of microglial activation in multiple sclerosis patients using positron emission tomography Visualization and genetic modification of resident brain microglia using lentiviral vectors regulated by microRNA-9 Seven-tesla MRI and neuroimaging biomarkers for 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interactions-a new role for the tumor perivascular space as highway for trafficking microglia Two photon intravital microscopy of lyme borrelia in mice New tools for studying microglia in the mouse and human CNS In vivo imaging of activated microglia in a mouse model of focal cerebral ischemia by two-photon microscopy Dynamics of the microglial/amyloid interaction indicate a role in plaque maintenance Monitoring in vivo function of cortical microglia A new approach for ratiometric in vivo calcium imaging of microglia Postmortem MRI and histology demonstrate differential iron accumulation and cortical myelin organization in early- and late-onset Alzheimer’s disease Protein-targeted gadolinium-based magnetic resonance imaging (MRI) contrast agents: design and mechanism of action Growth arrest of individual senile plaques in a model of Alzheimer’s disease observed by in vivo multiphoton microscopy ATP mediates rapid microglial response to local brain injury in vivo Fibrinogen-induced 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Pro-inflammatory activation of primary microglia and macrophages increases 18 kDa translocator protein expression in rodents but not humans Brain inflammation accompanies amyloid in the majority of mild cognitive impairment cases due to Alzheimer’s disease Microglia promote learning-dependent synapse formation through brain-derived neurotrophic factor Diverse microglial motility behaviors during clearance of dead cells in hippocampal slices Antibody-mediated clearance of amyloid-β peptide from cerebral amyloid angiopathy revealed by quantitative in vivo imaging Evaluation of TgH(CX3CR1-EGFP) mice implanted with mCherry-GL261 cells as an in vivo model for morphometrical analysis of glioma-microglia interaction Microglia-specific targeting by novel capsid-modified AAV6 vectors Distinct myeloid cell subsets promote meningeal remodeling and vascular repair after mild traumatic brain injury Microglia sculpt postnatal neural circuits in an activity and complement-dependent manner 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in brain inflammation and injury TREM2 function in Alzheimer’s disease and neurodegeneration Clinical neuroimaging using 7 T MRI: challenges and prospects The effects of ecstasy on neurotransmitter systems: a review on the findings of molecular imaging studies In vivo two-photon microscopy of the hippocampus using glass plugs Imaging microglial activation during neuroinflammation and Alzheimer’s disease Advanced motion compensation methods for intravital optical microscopy Redox indicator mice stably expressing genetically encoded neuronal roGFP: versatile tools to decipher subcellular redox dynamics in neuropathophysiology Resting microglia directly monitor the functional state of synapses in vivo and determine the fate of ischemic terminals Two-photon microscopy of deep intravital tissues and its merits in clinical research TREM2-mediated early microglial response limits diffusion and toxicity of amyloid plaques Microglia remodel synapses by presynaptic trogocytosis and spine head filopodia induction Multi-photon microscopy in cardiovascular research Rapid cell death is preceded by amyloid plaque-mediated oxidative stress Fate mapping reveals origins and dynamics of monocytes and tissue macrophages under homeostasis Bacskai BJ and Garcia-Alloza M (2018) In Vivo Imaging of Microglia With Multiphoton Microscopy Received: 29 March 2018; Accepted: 26 June 2018; Published: 19 July 2018 Copyright © 2018 Hierro-Bujalance, Bacskai and Garcia-Alloza. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) distribution or reproduction in other forums is permitted provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited in accordance with accepted academic practice distribution or reproduction is permitted which does not comply with these terms *Correspondence: Monica Garcia-Alloza, bW9uaWNhLmdhcmNpYUB1Y2EuZXM= Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher 94% of researchers rate our articles as excellent or goodLearn more about the work of our research integrity team to safeguard the quality of each article we publish Jan 28 (EFE).- Chinese artificial intelligence startup DeepSeek has temporarily limited new user registrations after suffering “large-scale malicious attacks” just hours after triggering a sharp fall in US stocks “Due to large-scale malicious attacks on DeepSeek’s services we are temporarily limiting registrations to ensure continued service Thanks for your understanding and support,” the company said on its status page The platform is “investigating the issue” that has caused its web and application programming interface (API) services to operate with “degraded performance,” it added DeepSeek has not yet specified the nature or origin of the attacks launched in 2023 by a Chinese hedge fund and practically anonymous internationally until a few weeks ago who said its sudden success should be a “wake-up call” for the US tech sector Trump said he still expected American tech companies to master artificial intelligence but acknowledged the challenge posed by DeepSeek which was the most downloaded app on the Apple Store over the weekend “The release of DeepSeek AI from a Chinese company should be a wake-up call for our industry,” the US president said Monday The emergence of DeepSeek’s AI model caused the New York’s Nasdaq exchange to fall by more than 3 percent hitting the technology giant Nvidia (-16.9 percent) in particular which suffered the biggest one-day fall for a listed company in history Analysts say DeepSeek is a threat to the market because of its commitment to open source and its low costs: its developers claim that the latest model was trained for only 55 days with a budget of less than $6 million “Deepseek’s R1 is an impressive model particularly around what they’re able to deliver for the price We will obviously deliver much better models and also it’s legit invigorating to have a new competitor We will pull up some releases,” OpenAI CEO Sam Altman said on X the impact of DeepSeek was felt by other tech companies such as Broadcom (-17 percent) Alphabet (-4.2 percent) and Microsoft (-2.14 percent) as well as energy-related companies such as Constellation Energy (-21 percent) and Vistra (-30 percent) with Chinese stock markets closed for the Lunar New Year major Asian markets followed Wall Street’s lead and closed with losses led by Tokyo’s Nikkei (-1.39 percent) DeepSeek’s breakthrough came almost in parallel with the unveiling of the ‘Stargate Project’ in the US which aims to invest around $500 billion over the next four years to build up to 20 new data centers to support AI initiatives in the country Saint Thomas the Apostle: In the face of doubts The priest Eugenio Bujalance offers Exaudi readers this article about Saint Thomas the Apostle Today the Church celebrates the feast of Saint Thomas The name Thomas in Aramaic means “twin” and the nickname by which the apostle “Didymus” was known in Greek has the same meaning perhaps a fisherman and one of the first to leave everything to follow Jesus his remains are found in the Ortona church dedicated to him Many have tried to explain and understand why he is called by this nickname of the Twin and he owes his celebrity to his questions and his doubts When he returns to the place where the disciples were hidden the disciples tell him: “We have seen the Lord” (Jn 20 he says: “If I do not see in his hands the sign of the nails unless I put my finger in the hole of the nails and put my hand in his side That is why he will also go down in posterity Saint Thomas will also be the one who makes the most perfect profession of Faith when he says: “My Lord and my God” (Jn 20 I once read a meditation where he said that Thomas’s twin is each of us and in each of the challenges that we face every day It is not a sin to realize that some mysteries of our Faith are difficult to understand nor is it a sin to accept that our understanding cannot fully understand everything Newman said: “A thousand problems do not create a doubt” because questions if they are the result of an involuntary doubt are lawful as long as they are asked not to deny the truths of faith but to try to explain or understand them in order to then believe more and better It is somewhat unfair to pigeonhole this apostle due to the weakness of his faith at a specific moment in the Gospel of Saint John we see that he develops a questioning character At the Last Supper Jesus announces his departure Jesus answers: “I am the way and the truth and the life” (Jn 14:6) Thomas does not want to be left with anything that he does not understand he needs to know and understand what the path is that will take him to Jesus when Jesus prepares to leave for Bethany at the time of the death of Lazarus he is in danger and the disciples remind him: “Teacher recently the Jews were trying to stone you” (Jn 11:8) Thomas says to the other disciples: “Let us also go and die with him” (Jn 11:16) Thomas reminds us that following Jesus means living his life Thomas is associating himself with the very life of the Lord in his life there were more acts of faith than doubts Sometimes the same thing happens to us too we need to feel safe in what we see and touch Thomas’s tendency to disbelief is cured by touching the Lord it is the mercy of the Lord that is made available to us instead of responding according to our distrust allows himself to be touched and performs a miracle transforming the unbeliever into a believer Who can say that he truly loves Christ if he does not strive to know Him better how important it is to know and love what we believe in May this feast of Saint Thomas grant us this miracle in our lives let us thank the Lord because we are Believers as many as there are who want to and cannot it is good to entrust our doubts to a good spiritual director or a good priest in the confessional to care for and increase the formation that feeds our Faith May the Holy Spirit guide our faith at all times and make it grow so that we can come to firmly believe in the Lord like Saint Thomas en 1986 y ordenado sacerdote el 25 de junio 2016 Licenciado en Derecho Canónico por la Universidad de San Dámaso de Madrid Defensor del Vínculo del Tribunal Eclesiástico de Córdoba Colaborador del Secretariado Diocesano de Pastoral Universitaria y capellán de las Carmelitas Descalzas Reflection by Bishop Enrique Díaz: I will praise you Paris Marks 400th Anniversary of the Congregation of the Mission Reflection by Bishop Enrique Díaz: The Lord’s mercy is eternal Tweets by Pontifex Volume 13 - 2021 | https://doi.org/10.3389/fnagi.2021.741923 This article is part of the Research TopicProgress of Translational Medicine in Alzheimer's DiseaseView all 14 articles Alzheimer’s disease is the most common form of dementia and epidemiological studies support that type 2 diabetes (T2D) is a major contributor The relationship between both diseases and the fact that Alzheimer’s disease (AD) does not have a successful treatment support the study on antidiabetic drugs limiting or slowing down brain complications in AD is currently being tested in patients with AD in the Evaluating Liraglutide in Alzheimer’s Disease (ELAD) clinical trial the effects of LRGT on brain pathology when AD and T2D coexist have not been assessed We have administered LRGT (500 μg/kg/day) to a mixed murine model of AD and T2D (APP/PS1xdb/db mice) for 20 weeks We have evaluated metabolic parameters as well as the effects of LRGT on learning and memory Postmortem analysis included assessment of brain amyloid-β and tau pathologies as well as insulin and insulin-like growth factor 1 receptors LRGT treatment reduced glucose levels in diabetic mice (db/db and APP/PS1xdb/db) after 4 weeks of treatment LRGT also helped to maintain insulin levels after 8 weeks of treatment While we did not detect any effects on cortical insulin or insulin-like growth factor 1 receptor m-RNA levels LRGT significantly reduced brain atrophy in the db/db and APP/PS1xdb/db mice LRGT treatment also rescued neuron density in the APP/PS1xdb/db mice in the proximity (p = 0.008) far from amyloid plaques (p < 0.001) LRGT reduced amyloid plaque burden in the APP/PS1 animals (p < 0.001) as well as Aβ aggregates levels (p = 0.046) and tau hyperphosphorylation (p = 0.009) in the APP/PS1xdb/db mice Spontaneous bleeding was also ameliorated in the APP/PS1xdb/db animals (p = 0.012) and microglia burden was reduced in the proximity of amyloid plaques in the APP/PS1 and APP/PS1xdb/db mice (p < 0.001) while microglia was reduced in areas far from amyloid plaques in the db/db and APP/PS1xdb/db mice (p < 0.001) This overall improvement helped to rescue cognitive impairment in AD-T2D mice in the new object discrimination test (p < 0.001) and Morris water maze (p < 0.001) our data support the role of LRGT in reduction of associated brain complications when T2D and AD occur simultaneously as regularly observed in the clinical arena supporting a two-way cross-talk between both pathologies the mice received an initial dose of subcutaneous LRGT (25 μg/kg/day) that was increased to 50 and 500 μg/kg/day daily during the first week From day seven 500 μg/kg/day (133.3 nmol/Kg/day) of LRGT was administered to the mice daily for 20 weeks T2D debuts at ≈6 weeks of age in db/db mice and by 26 weeks of age both AD and T2D are fully established in APP/PS1xdb/db mice Therefore LRGT treatment commenced at 6 weeks of age an continued up to 26 weeks of age Mice that did not receive LRGT received daily subcutaneous filtered PBS (vehicle) In vivo experiments included 31–34 females and 37–40 males randomly assigned to the treatment to complete the groups (Control n = 9–10 Postmortem studies included 9–31 males and 11–26 females to complete the experimental groups (Control n = 3–8 All experimental procedures were approved by the Animal Care and Use Committee of the University of Cadiz in accordance with the Guidelines for Care and Use of Experimental Animals (European Commission Directive 2010/63/UE and Spanish Royal Decree 53/2013) and plasma insulin levels were determined before the commencement of the treatment and every 4 weeks until sacrifice at 26 weeks Blood glucose levels were measured from nicked tails using the glucometer Optium Xceed (Abbott blood was collected from the tail vein and placed in tubes with potassium-EDTA (Sarstedt and plasma fraction was stored at –80°C until it was processed Plasma insulin levels were measured using an ultrasensitive mouse enzyme-linked immunosorbent assay (ELISA) (Mercodia Inc. Behavioral assessment commenced after 18 weeks of treatment with LRGT The acquisition phase was assessed 12 days prior to sacrifice The pool was a round tank of 90 cm in diameter and water temperature was 21 ± 2°C The animals performed four trials/day for 4 days Swimming commenced in each of the four virtual quadrants that the pool was divided in The retention phase started 24 h after the acquisition phase was completed and it consisted of a single trial with the platform removed Time required to locate the platform in the acquisition phase percentage of time spent in quadrant 2 during the retention phase and swim speed were analyzed using the Smart software (Panlab “What” was defined as the difference in time exploring familiar and recent objects “where” was defined as the difference in time exploring displaced and non-displaced objects and “when” was defined as the difference between time exploring familiar non-displaced objects and time exploring recent non-displaced objects Motor coordination was assessed by the rotarod (Panlab Harvard Apparatus An animal was placed on a horizontal rod (3 cm in diameter and 5.7 cm wide) which is rotated around its longitudinal axis and the animal must walk forward to remain upright and not fall off (the height to fall is 16 cm) The animals were placed on the rod for 4 min at 4 revolutions per minute (rpm) for training purposes the speed was increased from 4 to 40 rpm within 1 min The time spent on the rod and the velocity when the animals fall was recorded the sections were dehydrated in 70% ethanol for 15 min and then incubated in a cresyl violet (Sigma tissue was fixed in 0.25% acetic acid in ethanol for 5 min and subsequently in 100% ethanol and xylene for 2 min Images were acquired with an optical Olympus Bx60 microscope with an Olympus DP71 camera Adobe Photoshop Elements and Image J software were used to process the images and measure cortex and hippocampus sizes Sections contiguous to those used for cresyl violet staining were incubated by Prussian blue iron staining and neutral red counterstaining as described previously (Desestret et al., 2009) Images were acquired with an Olympus Bx60 (Olympus Japan) microscope with an Olympus DP71 camera (Olympus Japan) to assess the complete cortex and hippocampus The images were analyzed using the Image J software to quantify hemorrhage burden in the cortex and hippocampus The Image J software was used to analyze the number and burden of Aβ deposits in the cortex and hippocampus Soluble and insoluble Aβ40 and Aβ42 were quantified in the cortex and hippocampus with colorimetric ELISA kits (Wako, Osaka, Japan) (Aβ40, Ref. 294-62501; Aβ42, Ref. 290-62601) as described, with minor modifications (Infante-Garcia et al., 2017) Tissue (5–10 mg) was homogenized in 50 μl of PierceTM IP Lysis Buffer (Thermo Fisher Scientific 87788) with HaltTM (Thermo Fisher Scientific 78440) phosphatase and protease inhibitor cocktail and centrifuged (14,500 rpm) for 12 min at 4°C Soluble Aβ40 and Aβ42 levels were measured in supernatants The resultant pellet was extracted with 50 μl of 70% formic acid and centrifuged at 14,500 rpm for 10 min Insoluble fraction was neutralized and diluted 1:30 with 1M Tris (pH 11) Human Aβ40 and Aβ42 provided in the kit were used for standard curves Aβ aggregates were quantified using Human Amyloid β (82E1-specific) Aβ Oligomers Assay Kit (IBL The cortex was homogenized (1/5 w/v) in Tris-buffered saline (TBS; 20 mM Tris-HCl pH 7.5) containing HaltTM phosphatase and protease inhibitor cocktail Homogenates were then centrifuged (14,500 rpm) for 60 min at 4°C The supernatant was collected and diluted 1:2 with EIA buffer provided in the kit All absorbances were measured spectrophotometrically at 450 nm (MQX200R2; Biotek Instruments Total tau and tau phosphorylation levels were measured in cortical and hippocampal samples as described previously (Infante-Garcia et al., 2017) Tissue was homogenized in PierceTM IP Lysis Buffer (Thermo Fisher Scientific 78440) phosphatase and protease inhibitor cocktail The homogenates were sonicated and centrifuged at 4°C for 5 min at 15,000 g and protein concentration was determined by Bradford protein assay (Bio-Rad Proteins were separated on 10% acrylamide-bisacrylamide gels followed by electrophoretic transfer to PVDF membranes (Bio-Rad Membranes were then immersed in blocking buffer (Thermo Fisher Scientific WB7050) for 1 h and incubated overnight at 4°C with mouse anti-phospho-tau antibody (1:1,000) (clon AT8) Thermo Fisher Scientific Membranes were washed and then incubated with Secondary Antibody Solution Alk-Phos Conjugated (Anti-Mouse) (Thermo Fisher Scientific and a chemiluminescent inmunodetection system for mouse and rabbit primary antibodies (Invitrogen and signal was detected using Novex AP Chemiluminescent Substrate (Thermo Fisher Scientific the membranes were incubated with anti-total tau (1:1,000) (DAKO Optical density was semi-quantified after normalizing to β-actin (Sigma A5441) (1:1,000) using the Image J software Phospho-tau/total tau ratios were represented as percentage of control values RNA was isolated from the cortex using TRIzolTM (Invitrogen 15596026) following the instructions of the manufacturer and resuspended in purified nuclease-free water The RNA was quantified using a BioTek SynergyTM Mx (BioTek Instruments Complementary DNA (cDNA) was obtained from 500-ng RNA using iScriptTM cDNA Synthesis Kit (Bio-Rad Laboratories Inc. USA) (Ref.1708890) on Techne Genius Thermal Cycler (Techne Ltd. The 15-μl RT-qPCR reaction mix contained 7.5 μl 2X iTaqTM Universal SYBR® Green Supermix (Bio-Rad Laboratories Inc. and rRNA18S) or 900 nmol (for IGF-I) of forward and reverse primers The PCR thermal profile included 40 cycles of denaturation at 95°C for 10 s annealing at temperature according to each set of primers (61°C for IR-varA and 6–9 mice per group were included in the study The mRNA level of rRNA18S was used as internal control Relative quantification values of mRNA expression were calculated as 2–ΔΔCt with the comparative Ct method internal control Ct values were subtracted from the gene-of-interest Ct values to derive a ΔCT value The relative expression of the gene of interest was then evaluated using the expression 2−ΔΔCt where the value for ΔΔCt was obtained by subtracting the ΔCt of the calibrator from each ΔCT using the mean of the control (Control animals) as the calibrator The oligonucleotide primers used in this study were designed by BLAST and were obtained from Merck KGaA (Madrid Spain) for IR-varA (FW:TTTGTCCCCAGGCCATCC-RV:ATCTGGAAGTGTGAGTGTGG) IR-varB (FW:AATGGTG CCGAGGACAGTA-RV:ATCTGGAAGTGTGAGTGTGG) and IGF-I (FW:CACAACTACTGCTCCAAAGACAAA-RV:TTTTC CGTCACCTCCTCCAC); rRNA18S (FW:CTCAACACGGGAA ACCTCAC-RV:CTCAACACGGGAAACCTCAC) followed by Tukey’s b or Tamhane test was performed when all the eight groups under study were compared One-way ANOVA was also performed when only treated and untreated APP/PS1 and APP/PS1xdb/db mice were analyzed (Aβ levels and neuronal curvature close to amyloid plaques) Two-way ANOVA (groupXday) was performed to analyze the acquisition phase in the MWM test The SPSS v.24 software was used for all the statistical analyses One-way ANOVA for independent samples was performed for further analysis of individual days during acquisition in the MWM When we analyzed glucose level evolution along treatment, we did not detect a significant groupXweek effect by two-way ANOVA for independent samples [F(35, 380) = 1.18, p = 0.227]. However, further assessment of individual days revealed that the LRGT treatment reduced glucose levels in diabetic mice (Figure 1A) plasma glucose in the db/db mice and APP/PS1xdb/db crosses were significantly increased compared with the Control and APP/PS1 mice This increase in non-fasting glucose levels was more severe in the APP/PS1xdb/db animals (p < 0.001) glucose levels were significantly higher in the untreated db/db and APP/PS1xdb/db mice when compared with the LRGT-treated and untreated Control and APP/PS1xdb/db mice (p < 0.01) Differences in glucose levels between the untreated diabetic (db/db and APP/PS1xdb/db mice) and non-diabetic mice were maintained up to week 26 (p < 0.01) The LRGT treatment helped in limiting hyperglucemia in the db/db mice and glucose levels reached control values by week 10 (after 4 weeks of LRGT treatment) LRGT reduced non-fasting glucose levels in the APP/PS1xdb/db mice when compared with the untreated APP/SP1xdb/db animals by 10 weeks of age and a similar profile was observed by 14 weeks of age glucose levels in the APP/PS1xdb/db-LRGT mice were similar to those detected in the Control and APP/PS1 animals Glycemic control was maintained in the db/db-LRGT and APP/PS1xdb/db-LRGT mice until the end of treatment (26 weeks of age) Figure 1. Long-term liraglutide (LRTG) treatment ameliorates metabolic alterations in T2D and AD-T2D mice. (A) Non-fasting plasma glucose, (B) insulin, and (C) body weight were measured every 4 weeks in controls, APP/PS1, db/db and APP/PS1xdb/db mice treated with vehicle or LGRT 500 μg/kg for 20 weeks (from weeks 6 to 26) (complete statistical analysis included as a Supplementary Material) (A) LRGT significantly reduced postprandial glucose levels in diabetic mice: (##p < 0.01 Control Control; ††p < 0.01 Control and APP/PS1xdb/db-LRGT; p < 0.001 vs (B) Insulin levels were maintained by long-term LRGT treatment immediately before the commencement of LRGT treatment insulin levels were significantly increased in APP/PS1xdb/db mice (##p < 0.01 Control (C) LRGT maintained body weight in APP/PS1-LRGT (##p < 0.01 vs Data are representative of 5–12 animals and differences were detected by one-way ANOVA followed by Tukey’s b or Tamhane test When we analyzed insulin levels, we detected a significant groupXweek effect by two-way ANOVA for independent samples along treatment [F(35, 384) = 2.64, p < 0.01]. Further differences were observed among the groups when we analyzed individual weeks (Figure 1B) basal insulin levels in untreated animals were increased in the db/db mice although differences with the Control and APP/PS1 animals only reached statistical significance in the APP/PS1xdb/db mice (p < 0.01) The fact that insulin levels were significantly increased in the APP/PS1xdb/db mice suggests an earlier metabolic compromise in the crossed model that requires an increase in pancreatic activity statistical differences were observed in the db/db-LRGT and APP/PSxdb/db-LRGT mice when compared with the non-diabetic animals (Control and APP/PS1) suggesting an increase in insulin production in the LRGT-treated mice that allows for better glycemic control (p < 0.01) This situation was maintained from 14 to 26 weeks of age and LRGT helped to increase insulin levels in the diabetic mice (db/db-LRGT and APP/PS1xb/db-LRGT) to control hyperglycemia LRGT helped in maintaining the body weight of the APP/PS1xd/db mice avoiding weight loss from week 22 until the end of the study (26 weeks) As described previously, episodic memory was affected in the APP/PS1, db/db, and APP/PS1xdb/db mice in the new object discrimination test (Ramos-Rodriguez et al., 2015; Infante-Garcia et al., 2016), reaching statistical significance in the case of “what” and “where” paradigms when all the groups under study were compared (Figure 2A) Differences reached statistical significance when the untreated APP/PS1xdb/db mice were compared in the “what” and “where” paradigms (p < 0.01) the LRGT treatment counterbalanced this situation and the APP/PS1xdb/db-LRGT mice performed like the Control mice in both paradigms no differences were observed in any of the paradigms (distance traveled in the open field and rotarod) when the db/db-LRGT or the APP/PS1xdb/db-LRGT mice were compared with the untreated db/db or the APP/PS1xdb/db animals suggesting that the observed improvement in learning and memory is not due to changes in motor activity Motor activity assessment on liraglutide (LRGT)-treated mice Data are representative of five animals per group (308–920 neurons/group) (F) Illustrative examples of SMI-312 (red) and TS (green) staining in the proximity of and far from amyloid plaques (yellow lines mark representative neurites) (A) LRGT treatment reduced amyloid plaque burden in the cortex from APP/PS1 mice (**p < 0.01 vs rest of the groups; ††p < 0.001 vs No differences were observed in the hippocampus LRGT treatment also reduced amyloid plaque size in the cortex from APP/PS1xdb/db mice (p < 0.01 vs APP/PS1-LRGT; **p < 0.01 vs (B) Illustrative image of TS (green) and 4G8 (red) staining of amyloid plaques in the cortex from all the groups studied (C) No differences were observed when soluble Aβ40 or Aβ42 levels were analyzed in the cortex Aβ aggregates were significantly reduced in APP/PS1xdb/db mice on LRGT treatment (†p = 0.046 vs No statistical differences were observed in the hippocampus for soluble Aβ40 or Aβ42 levels Insoluble Aβ40 (‡p = 0.026 vs APP/PS1) and Aβ42 (‡p = 0.011 vs APP/PS1) levels are reduced in APP/PS1xdb/db when compared with APP/PS1 animals and LRGT treatment contributes to further reductions in the cortex No differences were observed in the hippocampus for insoluble Aβ40 or Aβ42 Data are representative of 6–9 animals (D) Tau phosphorylation was reduced in the cortex after LRGT treatment (††p = 0.009 vs Differences did not reach statistical significance in the hippocampus Data are representative of 3–11 mice (E) Illustrative example of Western blot for phospho-tau and β-actin in the cortex from all the groups studied (A) LRGT treatment reduces hemorrhage burden in the cortex (†p = 0.012 vs A similar profile was observed when we analyzed cortical hemorrhage density (††p < 0.001 vs No differences were detected in the hippocampus when hemorrhage burden or density was analyzed Data are representative of 3–5 mice (489–1,012 hemorrhages/group) (B) Illustrative example of cortical hemorrhages stained with Prussian blue Green arrows point at individual hemorrhages (C) LRGT treatment reduced cortical microglia burden in APP/PS1 and APP/PS1xdb/db mice in the proximity of amyloid plaques (**p < 0.01 vs LRGT also reduced microglia burden in cortical amyloid plaque-free areas in diabetic mice (**p < 0.01 vs No statistical differences were observed in the hippocampus close or far from amyloid plaques Data are representative of five mice (cortex 572–748 ROIs/group; hippocampus 108–230 (D) Illustrative example of cortical immunostaining for Iba1 (microglia Zoom-in images of representative regions are marked by white squares and presented next to the original image (E) No differences were observed in the cortex when we analyzed IR-A Data are representative of 6–9 mice When we analyzed IR-A mRNA expression, we did not observe any differences in the cortex from any of the groups under study (Figure 5E). Similar outcomes were observed for IR-B or IGF-1R (Figure 5E) reinforcing a neuroprotective role for LRGT in AD and T2D our studies did not include functional analysis Given the relevance Aβ pathology in AD further studies would be required to fully characterize Aβ structures and to provide a full picture of changes observed when AD and T2D coexist ultimately supporting a beneficial role of LRGT at the vascular level Our results support a positive role for LRGT at central level when AD and T2D coexist, as usually observed in the clinic (Janson et al., 2004) and cognitive impairment are significantly ameliorated supporting the beneficial role of LRGT in the brain and clinical studies on patients with AD The raw data supporting the conclusions of this article will be made available by the authors without undue reservation upon reasonable request The animal study was reviewed and approved by Junta de Andalucia (Guidelines for Care and Use of Experimental Animals European Commission Directive 2010/63/UE and Spanish Royal Decree 53/2013) and the University of Cadiz Bioethics Committee SL-L: design and critical revision of the manuscript for intellectual content MG-A: study concept and design and drafting and critical revision of manuscript for intellectual content All authors provided critical feedback and helped shape the research Instituto de Investigacion Biomedica de la Provincia de Cadiz (INIBICA) MG-A: Agencia Estatal de Investigación Programa Estatal de Generación de Conocimiento y Fortalecimiento Científico y Tecnológico del Sistema de I + D + i y del Programa Estatal de I + D + i Orientada a los Retos de la Sociedad del Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020 (PID2020-115499RB-I0) Programa Estatal de I + D + I orientada a los Retos de la Sociedad (BFU 2016-75038-R) financed by the Agencia Estatal de Investigación (AEI) and the Fondo Europeo de Desarrollo Regional (FEDER) Subvención para la financiación de la Investigación y la Innovación Biomédica y en Ciencias de la Salud en el Marco de la Iniciativa Territorial Integrada 2014–2020 para la Provincia de Cádiz financed by the Fondo de Desarrollo Regional (FEDER) (PI-0008-2017) All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher We thank the Animal Facility of the University of Cadiz and its personnel for their help and support We thank Servicios Centrales de Investigacion in Biomedicina (SC-IBM) from Universidad de Cadiz for the resources and technical support The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fnagi.2021.741923/full#supplementary-material Diabetes is related to cerebral infarction but not to AD pathology in older persons Google 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This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) On Sunday, January 8, the extraordinary concert of the Epiphany Gala of the Orquesta Filarmónica de Málaga, took place in Malaga, Spain, featuring Octavio J. Peidró on the podium, accompanied by the voices of soprano Paula Ramírez Álvarez and tenor Vicente Bujalance Leal (read more) Our collaborator Reme Díaz attended the concert and leaves us this detailed summary Among the musical traditions that come with the New Year there are the family concerts around the Epiphany date which take advantage of the holidays to bring music to the public not used to these events And nothing better than starting the year (and finishing Christmas holidays) with a tribute to Walt Disney through a program with different soundtracks from the “dream factory” performed by the Orquesta Filarmónica de Málaga (OFM) and conducted on this special occasion by Maestro Octavio J in the main auditorium of the Malaga Trade Fair and Conference Center (FYCMA) a great venue for conferences and exhibitions but not so good for hosting such a musical event which was sold out with an audience of all ages (among which we could see many children dressed as their favorite Disney or Marvel characters) and the lighting of the auditorium were unworthy of the wonderful work of the orchestra and the soloists the soprano Paula Ramírez Álvarez and the tenor Vicente Bujalance Leal We missed being able to enjoy the surround sound of the orchestra especially at the beginning of the program where the first theme could barely be heard at the back of the auditorium The string instruments were almost canceled out by the brass section and percussion which shows that even if they weren’t very experienced in this type of concerts music excites us regardless of the age or the conditions of the venue Strictly following the order of the program the song Something There from Beauty and the Beast (with arrangements by the conductor although the fantastic voices of the soloists were almost completely masked by the sound of the orchestra at its peak moments we could enjoy the main theme from Pocahontas masterfully performed by soprano Paula Ramírez who gave the song (composed by Alan Menken and lyrics by Howard Ashman) her operatic style This part of the concert reminded us of the selection of songs from the past Hollywood in Vienna tribute to Alan Menken saving the differences in terms of the venue and the scenography Courage from the original soundtrack by Jerry Goldsmith) was then performed by the OFM though we would have loved to continue enjoying Paula’s voice singing Reflection It didn’t take long for Paula to return to the stage this time to thrill us with a moving interpretation of the theme God Help the Outcasts from the movie The Hunchback of Notre Dame A film whose soundtrack (composed by Alan Menken and arranged for the concert by Octavio J Peidró) is forgotten in many Disney tributes and which we were happy to have on the program of this concert twice since that almost at the end of it the duet Someday by Paula and Vicente was also performed This soprano-tenor duet took over halfway through the concert to return to The Beauty and the Beast with the main theme where they shone more as soloists than in the part in which they combined their voices perhaps due to the more operatic range of Paula and the “Operetta” style of Vicente It caught our attention that the program didn’t gather the themes of each movie having jumps in their chronological order and even mixing animated films with live-action films from different periods and genres Famous themes from movies such as Frozen or Tangled were missing as well we were very excited to get the printed program we could be able to consult the pieces and the information of the performers and keep a lovely memory of the concert after a second duet from The Beauty and the Beast the orchestra gave us one of the best musical moments of the evening with the performance of a wonderful symphonic suite from The Chronicles of Narnia and where for the first time we noticed that the orchestra was able to shine brightly and where Maestro Octavio J Peidró transmitted his energy with a vigorous and passionate conduction that made him even jump off the stage several times The long-standing ovation with which the public responded was a good example of this we returned to the animation of the 90s with Vicente Bujalance singing Go the Distance from the film Hercules and we thought his powerful voice and lung capacity was on a level comparable to an elite athlete the song Part of Your Word from The Little Mermaid was one of the most emotional moments of the night hand in hand of course with soprano Paula Ramírez soprano and tenor combined acting and singing with their wonderful voices and a lovely connection between them and the orchestra for the classic theme from Mary Poppins arranged by Maestro Peidró from the very popular soundtrack composed by the Sherman brothers The program closed with a demanding suite from Pirates of the Caribbean where the orchestra brilliantly performed the different themes by Klaus Badelt and where the director surprised us again with his ability to jump on the stage and his movements full of energy and passion The audience answered with a standing ovation and Maestro Peidró thanked and greeted every member of the orchestra We thought it was an excellent way to end the concert before “boarding” a new year they surprised us with a suite that combined animation from the classic era such as the theme I wanna be like you from The Jungle Book with the soundtrack of Aladdin and the song Friend Like Me Mary Poppins returned to cheer us up with the songs A Spoonful of Sugar and Supercalifragilísticoexpialidocious where the audience sang and applauded encouraged by the conductor and amused by the sound of the orchestra the concert came to an end after a long final ovation from the attendees Despite the venue and the fact that the next day we returned to school or our work routines for a couple of hours we were able to move to a magical world where dreams are the protagonists A place where we were able to ask the Genie of the lamp that this new year should give us back the joy and illusion of when we were children Discover a way to enjoy that music live in SoundTrackFest Here you will find all the information you need to live your favorite Soundtracks and meet the Composers The Spanish Pediatric Association (Asociación Española de Pediatría AEP) has as one of its main objectives the dissemination of rigorous and up-to-date scientific information on the various areas of pediatrics Annals of Pediatrics is an open access journal that serves as the Association's Scientific Expression Organ and constitutes the vehicle through which members communicate as well as review articles prepared by experts in each specialty and guidelines or positioning documents prepared by the different Societies/Specialized Sections integrated into the Spanish Pediatric Association a reference for Spanish-speaking pediatrics is indexed in the most important international databases: Index Medicus/Medline IBECS the potential association between infection by these viruses and febrile seizures in healthy children has not been investigated in depth which led us to conduct a pilot study on the subject The study included children aged 6 months to 5 years without neurologic disease that presented with febrile seizures to the paediatric emergency department of a tertiary care hospital in Madrid between April 2019 and April 2020 a year when there was evidence of circulation of EV and HPeV based on data from the department of microbiology of the hospital Nasopharyngeal or rectal swabs were collected to perform polymerase chain reaction (PCR) tests for detection of EV and HPeV PCR testing for detection of these viruses was also performed in CSF We documented the development of convulsions in each patient after the initial visit to the emergency department for a 6-month follow-up period after the end of the recruitment period The statistical analysis was performed with the software IBM® SPSS® Statistics 20 We analysed data for a total of 39 patients, and the PCR test was positive for EV or HPeV in 15 (38.5%; 13/39 for EV and 2/39 for HPeV). Fig. 1 and Table 1 summarise the main results of the study treatment and outcomes and the 39 patients with febrile seizures by study group This may suggest that seizures may have to do with an individual predisposition rather than a specific infection it does not appear that the development of convulsive seizures in the context of EV or HPV infection is associated with an increased risk of recurring seizures compared to other aetiologies and the results should be interpreted with caution All the patients had favourable outcomes without neurologic sequelae 38.5% of the cases of febrile seizures were associated with infection by EV or HPeV (33.3% and 5.1% so these viruses should be taken into account as potential triggers of febrile convulsive seizures The main limitation of our study is that it was conducted in a single centre and that it did not include every patient that met the inclusion criteria Another important limitation is that few patients underwent lumbar puncture (n = 7) especially in the group positive for EV (n = 1) which limits the validity of our findings in this regard the viruses were identified in nasopharyngeal and/or rectal swab samples so it was not possible to establish with certainty that they were the aetiological agents involved in the infection We consider that more studies with larger samples and longer follow-up are required to establish the course and recurrence of febrile seizures in patients with infection by EV and HPeV Please cite this article as: García Sánchez P, de Ceano-Vivas la Calle M, Romero Gómez MP, García Bujalance S, Calvo Rey C. Asociacion de crisis febriles e infección por enterovirus y parechovirus humano en urgencias pediátricas: estudio piloto. An Pediatr (Barc). 2022;96:457–459. These authors have contributed equally. Anales de Pediatría (English Edition) follows the Recommendations for the Conduct Editing and Publication of Scholarly Work in Medical Journals African women are redefining their role in society challenging stereotypes and breaking barriers We present a brief journey through the diversity challenges and triumphs of women in the world's most diverse continent Africa is home to a population of 1.4 billion people spread across hundreds of ethnic groups speaking more than a thousand languages The role of women in this diverse landscape is deeply influenced by the specific culture of each region Some African countries have made significant progress in terms of equality Namibia and Ethiopia are also countries governed by women  Meanwhile, women in other African countries such as Burundi, the Central African Republic and Niger, considered the poorest and least developed in the world, seem to be stuck in prehistoric times. Not to mention the areas of the Democratic Republic of Congo, South SudanMozambique where women are victims of forced displacement and sexual violence the gap between women in urban and rural areas continues to be enormous there is a growing incorporation of women into the labor force traditional practices persist that limit their opportunities African women play a fundamental role in the continent's development Despite facing challenges such as violence poverty and lack of access to all kinds of resources women in Africa demonstrate exceptional resilience and leadership every day They are central to the informal economy or agriculture which are the basis of many African economies being largely responsible for food production and marketing.  investment in women's education or leadership training programs have enabled many women to start businesses increase their independence and contribute to local development Education translates immediately into lower poverty improved community health and better education for their children and leadership programs are propelling them to lead social and political movements fighting for their rights and for greater social and political representation Subscribe to Omnes magazine and enjoy exclusive content for subscribers a special concert was held in Malaga (Spain) where composer and conductor Arturo Díez Boscovich led the Orquesta Filarmónica de Málaga (Philharmonic Orchestra of Malaga) to premiere two works of his creation: the first one dedicated to the Soho Theater in Malaga and the second one dedicated to Don Quixote Casares on guitar, with Pablo Bujalance responsible for the dramaturgy (read news) and offers us this special article exclusively for SoundTrackFest NOTE: SoundTrackFest would like to send a special thanks to Pablo Bujalance for his dedication to leave us a few words about the event When we talk about soundtracks we’re used to concerts with orchestra and the “musical adventure for narrator and symphony orchestra of Don Quixote” we enjoyed last Friday February the 14th 2020 at the Auditorium Edgar Neville (Diputación de Málaga) was an amazing exercise of creativity where the soundtrack and the story blew the imagination of an almost complete venue Despite the multiple references that cinema television or animation has provided us for this archetype the concert reversed that passive attitude of the spectator and converting them into an imaginary protagonist and companion of the “Knight of the sad figure” And everything thanks to one of those rare symbiosis not frequently seen gathering an extraordinary cast that overflowed talent on the stage: the masterful score and conducting of maestro Arturo Diez Boscovich the impeccable artistic performance of the Philharmonic Orchestra of Malaga and the acclaimed Malaga-born guitarist Daniel Casares the immeasurable acting of veteran voice actor Camilo García and the adapted dramaturgy from Cervantes´s masterpiece by the hand of the great journalist and writer Pablo Bujalance the first thing was not the text but the musical composition presented in 2004 by Boscovich for a contest in the Italian city of Città di Castello The requirement was to focus the work on Don Quixote and it ended winning the contest Arturo had the idea of recovering it for symphonic orchestra with a narrator In 2019 he finally decided to shape it and that was when he contacted the journalist to write the texts “I started from the scenes that were already composed and then added others from myself we adjusted everything to fit perfectly music and text Everything inspired by the original Cervantes the result has little to do with that Quixote of 2004” the concert started with another soundtrack premiere composed by Boscovich for the recently released Antonio Banderas project An equally powerful and intimate score that reminds maestro John Williams and honors Banderas’ own spirit of effort and dedication until his dream is fulfilled to achieve a dream defeated by the sanity of reality Camilo García was in charge of the narration and of performing the rest of the characters (Don Quixote The idea of having him there appeared at the beginning as Pablo Bujalance tells us: “OFM (Philharmonic Orchestra of Malaga) proposed him to collaborate and he loved the idea but he had never been with an orchestra on stage.” A privilege to see him act not only with the voice but with the expressiveness of his gestures If we closed our eyes we could sometimes imagine Harrison Ford Gerard Depardieu or even Christopher Lee’s Saruman in “The Lord of the Rings.” (NOTE: He’s the Spanish dubbing voice for those actors in the movies) it was impossible to close our eyes and miss a second of the passionate conducting of Arturo Díez Boscovich or the great interpretation of the OFM and Daniel Casares The texts were interspersed or sometimes overlapping with music to narrate with words and musical notes the journey of Don Quixote Don Quixote confuses a tavern with a grand castle the composition evoked the epic greatness of the hero the rough but faithful personality of Sancho the painful madness or the love of Dulcinea with references to film composers such as Miklós Rósza Basil Poledouris or Spanish classical music such as Falla or Rodrigo these references or tributes that are born of admiration become a unique and unequaled work that nothing would have to envy those great music masters This original experience culminated in a standing ovation from the audience and demonstrating that film music DNA of Arturo the OFM performed the “Malaga Soundtrack” that Boscovich composed in 2015 for the 18th edition of the “Malaga Festival Spanish Cinema” as commissioned by one of its sponsors An exciting and unique concert and a privilege to have such great artists gathered around the figure of Don Quixote. An idea at the height of the great dreams and ideals of the character immortalized by Cervantes, and that we hope it’s only the beginning since it has shown us that, in the theater and music scene, there is still much to say. Do you like music from films, games, and TV series? Discover a way to enjoy that music live in SoundTrackFest. Here you will find all the information you need to live your favorite Soundtracks and meet the Composers. Bangkok, Apr 1 (EFE).- With human features and a soft voice, presenter Natcha debuted Monday on Thai television and became the country’s first reporter created with artificial intelligence. Natcha made her debut for the News Alert program on the Nation TV channel, donning a blue suit and reading the headlines on political information. Natcha, identified by a banner as an “AI presenter,” will soon be accompanied by Nitchan, a second reporter also generated by artificial intelligence. “AI reporters will help support the work of our editorial department, allowing (human) reporters more time to focus on gathering and verifying information,” Nation TV CEO Apirawee Pichayadecha announced last week. The director of the Thai channel, part of the media conglomerate Nation Group, said the new AI reporters would also serve as ambassadors for the channel, one of the main networks in the country, and participate in other news events. “The most important thing is that AI has no language barriers and can therefore provide unlimited information to viewers without any restrictions,” Apirawee said. The use of presenters generated with artificial intelligence is booming, mainly in Asia. In 2018, Chinese state news agency Xinhua put Qiu Hao on the air and made him the world’s pioneering AI presenter. Other countries such as India, Bangladesh and Pakistan have since created presenters with AI in their main newscasts. EFE Volume 8 - 2020 | https://doi.org/10.3389/fcell.2020.571258 This article is part of the Research TopicMechanisms of Neuronal Recovery in the Central Nervous SystemView all 18 articles The germinal matrix-intraventricular hemorrhage (GM-IVH) is one of the most devastating complications of prematurity The short- and long-term neurodevelopmental consequences after severe GM-IVH are a major concern for neonatologists These kids are at high risk of psychomotor alterations and cerebral palsy; however Erythropoietin (EPO) has been previously used to treat several central nervous system complications due to its role in angiogenesis EPO is regularly used to reduce the number of transfusions in the preterm infant EPO crosses the blood-brain barrier and EPO receptors are expressed in the human brain throughout development we have administered intraventricular collagenase (Col) to P7 mice as a model of GM-IVH of the preterm infant we have characterized our animals in the short (14 days) and the long (70 days) term EPO treatment significantly limited brain atrophy and ventricle enlargement EPO also restored neuronal density and ameliorated dendritic spine loss inflammation and small vessel bleeding were also reduced resulting in the preservation of learning and memory abilities as a feasible peripheral marker of GM-IVH-induced damage our data support the positive effect of EPO treatment in our preclinical model of GM-IVH A set of mice (10–12/group) were sacrificed at P14 to analyze the early effects of EPO and a second set (10–12/group) were aged up to P70 to assess the long-term effects of EPO treatment Two weeks before sacrifice mice underwent behavioral assessment Animals that died before the experiments were completed were not included in subsequent analyses All animals were maintained in the Animal Facility of the University of Cadiz under 12 h light/dark cycles and controlled temperature (21 ± 2°C) with ad libitum access to food and water All experimental procedures were approved by the University of Cadiz Bioethical Committee (Guidelines for Care and Use of Experimental Animals European Commission Directive 2010/63/UE and Spanish Royal Decree 53/2013) Rotarod (Ugo Basile Srl; Varese, Italia) was used to evaluate motor skills as described (Segado-Arenas et al., 2018) Mice were placed in the cylinder 4 min at 4 rpm for habituation purposes During the test speed was progressively increased from 4 to 60 rpm and the time spent on the rotarod was recorded A round pool (0.95 m in diameter) was filled with water (21 ± 1°C) and four equal virtual quadrants were indicated with geometric cues located in the walls. Experiments were run as previously described (Infante-Garcia et al., 2017) The acquisition started starting 12 days before sacrifice and consisted of 4 trials/day for 4 days with the submerged platform in the virtual quadrant number 2 Time limit to locate the platform was 60 s/trial with an intertrial interval of 10 min When the animal did not find the platform it was manually placed on it for 10 s Retention 1 commenced 24 h after the finalization of the acquisition phase and retention 2 was run 72 h after the end of the acquisition phase Retention phases consisted in a single trial with the platform removed Time required to locate the platform (acquisition) percentage of time spent in quadrant 2 (retention) and swimming velocity were analyzed using Smart software We analyzed brain morphology in 6 sections 1 mm apart (from 1.5 to −3.5 mm from Bregma). Sections were dehydrated in 70% ethanol for 15 min. They were stained with cresyl violet (Sigma, St. Louis, MO, United States) solution (0.5% w/v) for 10 min, as previously described (Ramos-Rodriguez et al., 2017) Sections were then fixed sequentially in 0.25% acetic acid Analysis was conducted blind to the experimental conditions Images were acquired using an optical Olympus Bx60 microscope (Japan) with an attached Olympus DP71 camera and Cell F software (Olympus Cortex and lateral ventricle sizes were measured using Adobe Photoshop and Image J software Consecutive sections to those used for cresyl violet staining were used for Prussian blue iron staining and neutral red counterstaining to analyze the presence of hemorrhages (Infante-Garcia et al., 2015) Hemorrhage burden (% of area covered by hemorrhages) was analyzed in the SVZ (up to 20 μm from the lateral ventricles) the cortex and the hippocampus using ImageJ software We quantified phopho-tau, total tau, phospo-Akt and total Akt levels in the striatum (including the SVZ), cortex and hippocampus from all animals under study. Samples were prepared as previously described (Infante-Garcia et al., 2016) tissue was homogenized in lysis buffer (Cell Signaling United States) with a protease/phosphatase inhibitor cocktail (Sigma at 15000 g and 4 °C Bradford (Bio-Rad Germany) protein assay was used for protein concentration in supernatants followed by electrophoretic transfer to PVDF membranes (Schleicher & Schuell Antibodies used included: phospho-tau clone AT8 (1:1000 phospho-Akt (Ser473) (1:1000) (Cell signaling anti-total Akt antibody (1:2000) (cell signaling United States) and anti-total tau antibody (1:1000) (DAKO Optical density was semi-quantified after normalizing to β-actin (1:2,500,000) (Sigma Data were represented as percentage of Control values Plasma matrix metalloproteinase 9 (MMP9) and gelsolin levels were measured, as feasible markers of central damage, in the short (P14) and the long (P70) term. We used ELISA kits for MMP9 (R&D System Corp, MN, United States) and gelsolin (Cusabio Biotech Co., Wuhan, Hubei Province, China), following manufacturer’s indications as previously described (Segado-Arenas et al., 2018) followed by Tuckey b test or Tamhane tests as required No statistical differences were detected between Sham and Control groups therefore all animals were combined in a single Control group Two-way ANOVA was used to analyze the MWM test (groupXday) and neurites architecture (groupXradius) SPSS v.15 software was used for all statistical analysis EPO treatment significantly reduces GM-IVH cognitive impairment at P70 (A) Episodic memory is significantly impaired at P70 after inducing a GM-IVH and EPO10 and EPO20 treatment counterbalances this limitation for “what” (**p < 0.01 vs “where” (**p < 0.01 vs Control) and “when” (**p < 0.01 vs (B) Spatial memory was also impaired after Col lesions when analyzed in the acquisition phase of the Morris water maze while EPO treatments significantly reversed this situation on individual days (day 1: p = 0.496 (C) A similar profile was observed in the retention phase and EPO10 and EPO20 treatment significantly improved the time spent where the platform used to be located Control + EPO20 and Col + EPO10) and 72 h (‡p = 0.025 vs Control and Col + EPO20) after completing the acquisition phase (D) Motor skills were not affected in rotarod (p = 0.962) spontaneous motor activity (p = 0.917) and swimming velocity (p = 0.566) No statistical differences were observed when EPO10 and EPO20 treatments were compared and even though a slightly higher improvement was observed in the cortex from EPO10-treated animals at P70 an overall better response was observed after EPO20 treatments in all the other paradigms under study as well as hemisection and ventricle sizes at P14 and P70 EPO treatment limits brain atrophy and neuronal alterations after induction of a GM-IVH with Col (A) Ipsilateral hemisection size was reduced in P14 and P70 mice while EPO treatment limited this effect (P14: ‡‡p = 0.003 vs Control + EPO10; P70: *p = 0.017 vs Differences observed in the cortex did not reach statistical significance at P14 (p = 0.358) although at P70 cortical compromise was ameliorated by EPO treatment (‡p = 0.014 vs Significant ventricle enlargement was detected at P14 and EPO treatment reversed this effect (**p = 0.001 vs rest of the groups) and a similar profile was observed at P70 (††p = 0.001 vs (B) Illustrative example of cresyl violet staining showing the ipsilateral ventricle enlargement at P14 and P70 while ventricle size is maintained in mice treated with EPO NeuN/DAPI ratio was reduced in the ipsilateral cortex and EPO treatment improved this situation (#p = 0.034 vs A similar profile was observed at P70 (**p < 0.001 vs The SVZ was also affected after Col lesions and EPO treatment significantly improved the NeuN/DAPI ratio both at P14 and P70 (P14: ††p = 0.002 vs Control + EPO20 and Col + EPO20; P70: ††p = 0.002 vs (D) Illustrative example of NeuN (red)/DAPI (blue) staining in the ipsilateral cortex and SVZ from mice with GM-IVH induced by Col where a reduction in NeuN+cells can be observed while more NeuN+cells are detected after EPO10 and EPO20 treatment (E) Neuronal complexity was affected by Col lesions and EPO treatment significantly improved this effect at P14 (**p < 0.01 vs Cont + EPO20; ‡‡p = 0.003 vs Cont + EPO20; T̄T̄p < 0.01 vs rest Control and Cont + EPO20) or P70 (**p < 0.01 vs Control + EPO20; ††p < 0.01 vs (F) Neuron curvature ratio was also impaired after Col lesions and EPO treatment significantly ameliorated this situation both at P14 (**p < 0.01 vs rest of the groups) and P70 (**p < 0.01 vs (G) Spine density compromise was also limited after EPO treatment in the ipsilateral hemispheres at P14 (**p < 0.01 vs Control and Control + EPO10) and at P70 (**p < 0.01 vs (H) Illustrative example of spines labeled by Golgi-Cox staining A reduction in spine density is observed in mice after lesions with Col and this effect is limited by EPO10 and EPO20 treatment at P14 and P70 The hippocampus showed a similar trend (hemorrhage burden: P14 [F(5,124) = 2.79 rest of the groups]; P70 [F(5,103) = 13.01 ∗∗p < 0.01 vs Control + EPO10 and Control + EPO20]; number of hemorrhages: P14 [F(5,124) = 2.33 Control + EPO20 and Col + EPO10] and P70 [F(5,116) = 15.80 Col + EPO20] and hemorrhage size: P14 [F(5,285) = 3.42 EPO treatment reduces vascular damage and inflammation after inducing a GM-IVH (A) Hemorrhage burden was significantly increased in the SVZ after Col lesions and EPO10 and EP20 treatments reversed this effect both in the short (P14) and the long term (P70) (P14: *p = 0.013 vs rest of the groups and P70: **p < 0.01 vs The number of individual hemorrhages was reduced after EPO treatment (P14: *p = 0.021 vs rest of the groups; P70: **p < 0.01 vs while individual hemorrhage size was not affected (P14: p = 0.169]; P70: p = 0.440] When we analyzed the cortex we observed a similar profile and increased hemorrhage burden was reduced by EPO treatment (P14: **p < 0.01 vs due to a reduction in the number of individual hemorrhages after the treatment (P14: **p < 0.01 vs rest of the groups; P70: ††p < 0.01 vs while hemorrhage size was not affected (P14: p = 0.123; P70: p = 0.296) increased hemorrhage burden in the ipsilateral side was reversed by EPO administration (P14: *p = 0.02 vs The number of individual hemorrhages was reduced after the treatment (P14: †p = 0.046 vs Control + EPO20 and Col + EPO10; and P70: **p < 0.01 vs Col + EPO20) while hemorrhage size was not affected (P14: p = 0.005 no further differences detected; P70: p = 0.270) (B) Illustrative images of cortical Prussian blue staining and Congo red counterstain for hemorrhages Increased bleeding after Col lesions was reduced by EPO treatment both at P14 and P70 Green arrow point at individual cortical hemorrhages Bottom panels show increased hemorrhage burden in the SVZ from Col-lesioned animals (C) Microglia activation after Col lesions was reduced by EPO treatment at P14 and P70 in the SVZ (P14: ##p = 0.003 vs Control and Control + EPO20; P70: **p < 0.01 vs cortex (P14: ‡‡p < 0.01; P70: **p < 0.01 vs Control) and hippocampus (P14: ‡‡p < 0.01 vs Control + EPO10 and Control + EPO20; P70: p = 0.098) (D) Illustrative example of microglia immunostaining for IBA-1 (green) in the SVZ and cortex from all groups under study Increased microglia activation can be observed in Col-treated mice while EPO treatment significantly reduces this effect EPO treatment reduces tau and Akt phosphorylation after Col lesions (A) Increased tau phosphorylation after Col lesions was reduced after EPO treatment at P14 although differences did not reach statistical significance (striatum: p = 0.193; cortex: p = 0.922 and hippocampus: p = 0.504]) A similar profile was observed when we analyzed animals in the long term (P70) (striatum: #p = 0.023 vs Control + EPO20; cortex: p = 0.658 and hippocampus: p = 0.522) (B) Illustrative example of phospho-tau/total tau striatum westerns blots at P14 and P70 showing an increased phosphorylation after Col lesions at P70 (C) Increased phosphor-Akt/total Akt ratios after Col lesions were reduced by EPO treatment in the ipsilateral hemisphere although differences did not reach statistical significance at P14 (striatum: p = 0.127; cortex: p = 0.552; hippocampus: p = 0.055) or at P70 (striatum: p = 0.931; cortex: p = 0.425; hippocampus: p = 0.475) (D) Illustrative example of striatum phosphor-Akt/total Akt western blots at P14 and P70 Effect of EPO treatment on peripheral markers of GM-IVH (A) EPO treatment does not affect MMP9 plasma levels at P14 (p = 0.633) or P70 (p = 0.453) (B) EPO treatment restores plasma gelsolin levels after inducing a GM-IVH in the short term while differences are no longer detected in the long term (P70) (P14 **p = 0.002 vs we only observed a slight increase in phopho-Akt levels after inducing a GM-IVH EPO treatment significantly restores plasma gelsolin levels suggesting that its positive effects at central level are also detected in the periphery and reinforcing further studies on the utility of plasma gelsolin as a prognostic tool European Commission Directive 2010/63/UE and Spanish Royal Decree 53/2013) and the University of Cádiz Bioethics Committee IB-F: study concept and design and critical revision of manuscript for intellectual content and critical revision of manuscript for intellectual content All authors read and approved the final manuscript MG-A Programa Estatal de I+D+I Orientada a los Retos de la Sociedad (BFU 2016-75038-R) financed by the Agencia Estatal de Investigación (AEI) and Fondo Europeo de Desarrollo Regional (FEDER) I+D+i Programa operativo FEDER Andalucía 2014–2020 FEDER-UCA18-107189 MG-A and IB-F Proyectos de Investigación e Innovación para Grupos de Investigación INIBICA (LI19/11IN-CO34) Intraventricular hemorrhage in premature infants: mechanism of 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hemorrhage Lubian-Lopez S and Garcia-Alloza M (2020) Erythropoietin Improves Atrophy Bleeding and Cognition in the Newborn Intraventricular Hemorrhage Copyright © 2020 Hierro-Bujalance, Infante-Garcia, Sanchez-Sotano, del Marco, Casado-Revuelta, Mengual-Gonzalez, Lucena-Porras, Bernal-Martin, Benavente-Fernandez, Lubian-Lopez and Garcia-Alloza. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) *Correspondence: Simon Lubian-Lopez, c2ltb25wLmx1Ymlhbi5zc3BhQGp1bnRhZGVhbmRhbHVjaWEuZXM=; Monica Garcia-Alloza, bW9uaWNhLmdhcmNpYUB1Y2EuZXM= "Am I looking at you?" she asks before starting the interview And she immediately laughs as she realises how strange the question sounds coming from someone who is blind but I don't see you." Anyone or anything in front of her appears as a light since she lost her sight 20 years ago due to diabetes portraying a unique Lady Macbeth in a play she produced herself and which is about to receive the Ángeles Rubio-Argüelles Award at the Festival de Teatro de Málaga imagine if at the same time you have to count the steps from one place to another on the stage listening to the sound of footsteps on a dry leaf that marks the space and following the carpet on the floor that shows you the way Just "tricks" for her but superpowers for anyone else makes her debut at the Malaga event this Sunday 26 January (7pm admission 20 euros) with the premiere at the Teatro Echegaray of I a play written by Pablo Bujalance and directed by Lorena Roncero Inspired by true stories of homeless people the director has imagined a queen without a crown Her belongings fit in a dingy cart and her home is a bench in a park in Malaga "This Lady Macbeth has been on the run since she allegedly escaped from a tower and arrived here with nothing," says the director from Espacio La Amarilla The haughty 'lady' portrayed by Shakespeare is still there but there is also the fragile woman that Bujalance now discovers "She has had everything and now she has nothing At other times she has been very strong but now she is afraid and very lonely," reflects Zira Williams Her character constantly moves in this duality between the signs that remain of the great lady she was and the wounds of a woman for whom even everyday things And it is with this vulnerability that Esther Ruiz connects instantly "Much more than with the Lady Macbeth of the castle She is a strong woman who we are now taking to the terrain of fragility of her reinvention since her early twenties when blindness took her away from hairdressing for good She then began to do what she had always wanted to do but never could because of life's obligations even though she knew that at the time it would be much more difficult not giving up." This in spite of her mobility problems and her occasional shortness of breath because of asthma "Even if my legs are shaking at the end of the play because I can't do it any more I'm going to be here until they throw me out we also have difficult moments." And that's where the support network comes in because "there's always someone to give you a hand." As she speaks It's the first time they have worked together but it seems like they have done it all their lives The connection and affection between them is evident which she is working on with Zira Williams Producciones she has surrounded herself with a "great team." Pilar Esteban 'La Pili' Eskarnia and Alessandra García provide the voice-over for the witches The music of the play comes courtesy of Artesonao Miguel Almanza has made the "teaser." Elisa Postigo And Julián Navarro is her right-hand man in the production "I would love to have someone else with me Although there are many people who support us highlighting the reality of small-scale theatre production "It's that nowadays it's very difficult to start up a business and she has taken it upon herself with a courage that is to be admired," adds Lorena Roncero This is her second monologue and her second production (after her debut in 2021 with 'Manto') "but this one is much more complicated." The text is for her the "biggest challenge" "Then you get on the stage and you forget something It's about letting your intuition guide you" she explains For Lorena Roncero it is also a challenge and a "learning experience." "All the focus is on what she does on respecting the text as much as possible and on accompanying her so that she shines" the theatre festival will present her with the Ángeles Rubio-Argüelles Award after the performance which recognises the work of women on stage but what a beautiful thing I'm going to experience," she concludes Comentar es una ventaja exclusiva para registrados