Astrocytic A2A receptors silencing negatively impacts hippocampal synaptic plasticity and memory of adult mice.

Astrocytes are wired to bidirectionally communicate with neurons namely with synapses, thus shaping synaptic plasticity, which in the hippocampus is considered to underlie learning and memory. Adenosine A2A receptors (A2A R) are a potential candidate to modulate this bidirectional communication, since A2A R regulate synaptic plasticity and memory and also control key astrocytic functions. Nonetheless, little is known about the role of astrocytic A2A R in synaptic plasticity and hippocampal-dependent memory. Here, we investigated the impact of genetic silencing astrocytic A2A R on hippocampal synaptic plasticity and memory of adult mice. The genetic A2A R silencing in astrocytes was accomplished by a bilateral injection into the CA1 hippocampal area of a viral construct (AAV5-GFAP-GFP-Cre) that inactivate A2A R expression in astrocytes of male adult mice carrying "floxed" A2A R gene, as confirmed by A2A R binding assays. Astrocytic A2A R silencing alters astrocytic morphology, typified by an increment of astrocytic arbor complexity, and led to deficits in spatial reference memory and compromised hippocampal synaptic plasticity, typified by a reduction of LTP magnitude and a shift of synaptic long-term depression (LTD) toward LTP. These data indicate that astrocytic A2A R control astrocytic morphology and influence hippocampal synaptic plasticity and memory of adult mice in a manner different from neuronal A2A R.

Cost-effectiveness analysis of COVID-19 tests in the unified health system.

BACKGROUND: To evaluate the cost-effectiveness ratio and economic impact of the Rapid Antigen Test (TR-Ag) to replace RT-PCR for the detection of the new Coronavirus in the Unified Health System (SUS). METHODS: This is a cost-effectiveness analysis. Clinical protocols were used for the diagnosis of COVID-19 at the São José Municipal Hospital, located in the city of Itaberá-SP. The Incremental Cost-Effectiveness Ratio (ICER) was divided into two scenarios. In the first, the accuracy reported by the test manufacturers was included, and in the second, the cost resulting from a systematic review. Both were compared with the performance of the RT-PCR test. The increase in diagnoses was chosen as a health outcome and absenteeism was used as a criterion for assessing the economic impact. RESULTS: The analysis resulted in incremental cost-effectiveness ratios of R$ 42,136.67 and R$ 68,329.73 for every thousand tests, according to the accuracy of the manufacturers' TR-Ag tests and what is reported in the literature in relation to RT-PCR, respectively. The average value found for the RT-PCR test (R$ 202.87) represents an increase of 165.32% in cost in relation to the value found for the TR-Ag. 4,305 tests were performed between April 2020 and December 2021 at the referral hospital. Also, maintaining the use of RT-PCR as the first choice for diagnosing COVID-19 and regulating absenteeism in the economically active population could have an impact of up to R$ 1,022,779.68 on municipal management. CONCLUSION: It is concluded that the TR-Ag are configured as a cost-effective alternative for the SUS in the detection of the new Coronavirus. The strategy becomes economically favorable for the expansion of testing, combating the COVID-19 pandemic and reducing the impact on the local economy. However, studies are needed to validate the accuracy of the tests so that economic evaluations on the subject are more assertive.

Downregulation of Sirtuin 1 Does Not Account for the Impaired Long-Term Potentiation in the Prefrontal Cortex of Female APPswe/PS1dE9 Mice Modelling Alzheimer's Disease.

Alzheimer's disease (AD), which predominantly affects women, involves at its onset a metabolic deregulation associated with a synaptic failure. Here, we performed a behavioral, neurophysiological and neurochemical characterization of 9-month-old female APPswe/PS1dE9 (APP/PS1) mice as a model of early AD. These animals showed learning and memory deficits in the Morris water maze, increased thigmotaxis and anxiety-like behavior and showed signs of fear generalization. Long-term potentiation (LTP) was decreased in the prefrontal cortex (PFC), but not in the CA1 hippocampus or amygdala. This was associated with a decreased density of sirtuin-1 in cerebrocortical synaptosomes and a decreased density of sirtuin-1 and sestrin-2 in total cerebrocortical extracts, without alterations of sirtuin-3 levels or of synaptic markers (syntaxin, synaptophysin, SNAP25, PSD95). However, activation of sirtuin-1 did not affect or recover PFC-LTP deficit in APP/PS1 female mice; instead, inhibition of sirtuin-1 increased PFC-LTP magnitude. It is concluded that mood and memory dysfunction in 9-month-old female APP/PS1 mice is associated with a parallel decrease in synaptic plasticity and in synaptic sirtuin-1 levels in the prefrontal cortex, although sirtiun1 activation failed to restore abnormal cortical plasticity.

Cartoon diplomacy: visual strategies, imperial rivalries and the 1890 British Ultimatum to Portugal.

This paper offers a novel interpretation of the 1890 British Ultimatum, by bringing to the front of the stage its techno-diplomatic dimension, often invisible in the canonical diplomatic and military narratives. Furthermore, we use an unconventional historical source to grasp the British-Portuguese imperial conflict over the African hinterland via the building of railways: the cartoons of the politically committed and polyvalent Portuguese artist and journalist Rafael Bordalo Pinheiro (1846-1905), published in his journal Ponto nos iis, from the end of 1889 and throughout 1890. We argue that the Ponto nos iis cartoons played a so far overlooked role in the unfolding of British-Portuguese affairs, as they shaped at a distance a diplomatic exchange with the British satirical journal Punch. Attacking and counterattacking his fellow cartoonists in Britain, Pinheiro surged into the role of informal diplomat. This cartoon visual and public diplomacy unfolded in the pages of both journals and was tied to the two countries' colonial conquests in Africa, where the Portuguese and British empires were competing to dominate the African hinterland through large technological systems. Hence the cartoons made visible to wider audiences the otherwise hidden role that technologies played in the two countries' affairs. In turn, the cartoons aimed at persuading the Portuguese public and ruling classes alike that only regime change, from monarchic to republican, would restore the wounded Portuguese national pride.

Identification of the calpain-generated toxic fragment of ataxin-3 protein provides new avenues for therapy of Machado-Joseph disease| Spinocerebellar ataxia type 3.

AIMS: Machado-Joseph disease (MJD) is the most frequent dominantly inherited cerebellar ataxia worldwide. Expansion of a CAG trinucleotide in the MJD1 gene translates into a polyglutamine tract within ataxin-3, which upon proteolysis may lead to MJD. The aim of this work was to understand the in vivo contribution of calpain proteases to the pathogenesis of MJD. Therefore, we investigated (a) the calpain cleavage sites in ataxin-3 protein, (b) the most toxic ataxin-3 fragment generated by calpain cleavage and (c) whether targeting calpain cleavage sites of mutant ataxin-3 could be a therapeutic strategy for MJD. METHODS: We generated truncated and calpain-resistant constructs at the predicted cleavage sites of ataxin-3 using inverse PCR mutagenesis. Lentiviral vectors encoding these constructs were transduced in the adult mouse brain prior to western blot and immunohistochemical analysis 5 and 8 weeks later. RESULTS: We identified the putative calpain cleavage sites for both wild-type and mutant ataxin-3 proteins. The mutation of these sites eliminated the formation of the toxic fragments, namely, the 26-kDa fragment, the major contributor for striatal degeneration. Nonetheless, reducing the formation of both the 26- and 34-kDa fragments was required to preclude the intranuclear localisation of ataxin-3. A neuroprotective effect was observed upon mutagenesis of calpain cleavage sites within mutant ataxin-3 protein. CONCLUSIONS: These findings suggest that the calpain system should be considered a target for MJD therapy. The identified calpain cleavage sites will contribute to the design of targeted drugs and genome editing systems for those specific locations.

Effect of Helicobacter pylori Infection on Macromineral and Trace Element Status-Systematic Review and Meta-Analysis.

OBJECTIVES: To explore the effect of Helicobacter pylori (Hp ) infection on zinc, copper, calcium, magnesium, phosphorus, and iodine status in the pediatric population. METHODS: A protocol was registered on PROSPERO. A literature search was conducted on Embase, PubMed MEDLINE, and Web of Science, from inception to September 2020, including all studies in English, Spanish, and Portuguese languages. Reference lists were manually searched. Primary studies describing at least one micronutrient status in Hp -positive and Hp -negative or control children were included. PRISMA recommendations were applied. Pooled mean differences (MDs) and 95% confidence intervals were estimated using a random-effects model. A total of 1011 citations were screened. Six cross- sectional studies were included. No publications regarding phosphorus and iodine were identified. RESULTS: Included studies in meta-analyses comprised an overall age range of 4-18 years, with Hp positivity ranging between 29.5% and 72.3%. These meta-analyses demonstrated a lack of evidence of an association between Hp -positive and Hp -negative or control children regarding serum zinc (vs Hp -negative: MD -1.36 µg/dL; vs control: MD 326.22 µg/dL), copper (MD -0.83 µg/dL), and calcium (MD 0.09 mg/dL) status. Considerable heterogeneity was recognized, except for calcium analysis (I 2 = 0%). Meta-analysis for magnesium was not performed. Five studies presented a low risk of bias. CONCLUSIONS: The study demonstrated a lack of evidence of an effect of Hp infection on serum zinc, copper, and calcium status. Studies concerning magnesium, phosphorus, and iodine status are warranted. Furthermore, larger and well-controlled studies are recommended.

First Report of Mulberry Rust Caused by Cerotelium fici on Morus nigra in Brazil.

Leaf rust caused by Cerotelium fici (Cast.) Arth. is the main disease affecting Moraceae family plants, such as Ficus and Morus species (Galleti and Rezende 2016; Srikantaswamy et al. 2006). In August 2020, rust symptoms were observed in 100% of mulberry (Morus nigra L.) trees in an experimental orchard (Piracicaba, SP, Brazil; 22°42'28"S, 47°37'42"W). Mulberry leaves with high rust severity became yellowish and fell-off prematurely. Pustules were light brown with yellowish halo and presented mean size of 0.9 mm2. Uredinial paraphyses (n = 50) measured 42.2 ± 0.67 µm long with wall uniformly ca 0.6-1.1 µm thick. Urediniospores were brownish, echinulate, globoid to broadly ellipsoid, and measured 27.1 ± 0.29 × 21.0 ± 0.27 µm with a wall thickness of 0.6 ± 0.01 µm (n = 100). The morphology of the urediniospores observed in this study was similar to that reported in the literature for C. fici on Morus alba and Ficus spp. (Gupta et al. 1994; McKenzie 1986; Hennen et al. 2005). We used a low-coverage genome-skimming approach to retrieve genetic information of the rRNA cluster and the mtDNA. Genomic DNA was extracted from 3-4 mg of stored urediniospores at -80 °C, macerated in liquid nitrogen, using a modified cetyl trimethylammonium bromide extraction procedure (Lo Piccolo et al. 2012), and sequenced with 150-bp paired-end reads on Illumina NovaSeq 6000 System. Raw data, (45,761,957 X 2 reads) were assembled with SPAdes v3.15.1 (Bankevich et al., 2012) and the output used to create a custom BLAST database. Loci used for the phylogenetic analyses were identified by BLASTn using, as a query, sequences of C. fici from Ficus sp. from Australia publicly available: Accession No. MH047210.1 for the rRNA and MW036502.1 for COX3. The retrieved sequences were deposited in GenBank under accession numbers OM296992 and OP797407 for the partial rRNA cluster and COX3, respectively. The Bayesian inference phylogenetic analysis of the three concatenate loci (18S, 28S, and COX3) revealed that the isolate obtained in this study (MN1) was clustered in a well-supported clade with C. fici type species. Pathogenicity tests were conducted using mulberry potted plants under greenhouse conditions (25 ± 5 °C). The urediniospores suspension (5 × 104 urediniospores ml-1) with 0.05% Tween 20 was sprayed with an airbrush on fully expanded leaves until run-off. As a control, mulberry plants were sprayed with distilled water and kept under the same conditions. Inoculated and mock-inoculated plants were kept in a dark moist chamber at 23 °C (± 2 °C) for 24 h. After this period, plants were moved to the greenhouse. The experimental design was completely randomized with five replicates, each replicate consisted of one potted plant and the experiment was performed twice. At 12 days post-inoculation, all inoculated plants showed rust symptoms identical to those observed in the field, whereas control plants had no symptoms. The first symptoms were small pustules on the abaxial surface of fully expanded leaves. Small chlorotic lesions were observed on the adaxial leaf surface, which evolved into necrotic lesions. The pathogen was re-inoculated into potted plants, where it was maintained through monthly inoculations. To our knowledge, this is the first report of mulberry rust on M. nigra in Brazil. As mulberry leaves are the only natural food for silkworm (Bombyx mori L.), rust poses a significant threat to the sericulture industry because the disease can decrease production and quality of mulberry foliage.

CD73-Mediated Formation of Extracellular Adenosine Is Responsible for Adenosine A2A Receptor-Mediated Control of Fear Memory and Amygdala Plasticity.

Adenosine A2A receptors (A2AR) control fear memory and the underlying processes of synaptic plasticity in the amygdala. In other brain regions, A2AR activation is ensured by ATP-derived extracellular adenosine formed by ecto-5'-nucleotidase or CD73. We now tested whether CD73 is also responsible to provide for the activation of A2AR in controlling fear memory and amygdala long-term potentiation (LTP). The bilateral intracerebroventricular injection of the CD73 inhibitor αß-methylene ADP (AOPCP, 1 nmol/ventricle/day) phenocopied the effect of the A2AR blockade by decreasing the expression of fear memory, an effect disappearing in CD73-knockout (KO) mice and in forebrain neuronal A2AR-KO mice. In the presence of PPADS (20 µM) to eliminate any modification of ATP/ADP-mediated P2 receptor effects, both AOPCP (100 µM) and the A2AR antagonist, SCH58261 (50 nM), decreased LTP magnitude in synapses of projection from the external capsula into the lateral amygdala, an effect eliminated in slices from both forebrain neuronal A2AR-KO mice and CD73-KO mice. These data indicate a key role of CD73 in the process of A2AR-mediated control of fear memory and underlying synaptic plasticity processes in the amygdala, paving the way to envisage CD73 as a new therapeutic target to interfere with abnormal fear-like emotional processing.

Growth of Methylobacterium organophilum in Methanol for the Simultaneous Production of Single-Cell Protein and Metabolites of Interest.

Research background: This study aims to monitor the growth of the methylotrophic bacteria Methylobacterium organophilum in a culture medium with methanol as a carbon source and to verify the production of unicellular proteins and other biomolecules, such as carotenoids, exopolysaccharides and polyhydroxyalkanoates, making them more attractive as animal feed. Experimental approach: Bacterial growth was studied in shake flasks using different carbon/nitrogen (C:N) ratios to determine their best ratio for achieving the highest volumetric productivity of cells and substrate consumption rate. This optimal parameter was further used in a fed-batch operating bioreactor system to define the kinetic profile of cell growth. Methanol consumption was measured by HPLC analysis and the extracted pigments were analyzed by liquid chromatography/mass spectrometry. Chemical composition and rheological properties of the produced exopolysaccharides were also determined. Results and conclusions: The best experimental parameters were verified using an initial methanol concentration of 7 g/L in the culture medium. The same initial substrate concentration was used in the fed-batch operation and after 60 h of cultivation 5 g/L of biomass were obtained. The accumulation of carotenoids associated with cell growth was monitored, reaching a concentration of 1.6 mg/L at the end of the process. These pigments were then analyzed and characterized as a set of xanthophylls (oxidized carotenoids). In addition, two other product types were identified during the fed-batch operation: exopolysaccharides, which reached a concentration of 8.9 g/L at the end of the cultivation, and an intracellular granular structure that was detected by transmission electron microscopy (TEM), suggesting the accumulation of polyhydroxyalkanoate (PHA), most likely polyhydroxybutyrate. Novelty and scientific contribution: Methylobacterium organophilum demonstrated a unique ability to produce compounds of commercial interest. The distinct metabolic diversity of this bacterium makes room for its use in biorefineries.

Economic analysis of the different endodontic instrumentation techniques used in the Unified Health System.

BACKGROUND: To assess the financial impact of incorporating a new (reciprocal) technology into endodontic treatments in the public health system (SUS). METHODS: This was a economic evaluation study (comparing the 3 different endodontic instrumentation techniques-manual, rotary and reciprocating), allocative efficiency analysis to optimize existing resources in the SUS, and financial contribution impact analysis of incorporation of a new technology. Thirty-one (31) 12 years-old volunteers were evaluated. RESULTS: The incremental cost-effectiveness ratio (ICER) was calculated at R$1.34/min, - R$0.60/min and BRL 0.10/min for the single-rooted, bi-rooted and tri-rooted teeth, respectively, when the rotary technique was compared with the manual type. In turn, the ICER was R$ 21.04/min, - R$ 0.73/min and - R$ 2.81/min for the 3 types of teeth, respectively, when the reciprocating technique was compared with the manual type. The incremental financial impact of replacing manual endodontic with rotary endodontic treatments would be - R$ 2060963.66 in the case of single-rooted teeth, but the number of treatments would also be reduced (- 19,379). In the case of two-rooted teeth, the incremental financial impact would be BRL 34921540.62 with the possibility of performing an additional 204,110 treatments. In turn, BRL 11523561.50 represented the incremental financial impact for teeth with 3 or more roots and with an increase of 72,545 procedures. When we analyzed the incremental financial impact of replacing manual endodontic with reciprocating endodontic treatments, it would be - R$ 730227.80 in the case of single-rooted teeth, allowing for an additional 2538 treatments. In turn, R$ 21674853.00 represented the incremental financial impact for bi-radicular teeth, with an increase of 121,700 procedures. In the case of two-rooted teeth, the incremental financial impact would be BRL 13591742.90 with the possibility of performing an additional 40,670 treatments. CONCLUSIONS: The reciprocating technique could improve access to endodontic treatment in the SUS as it allowed a simultaneous reduction in clinical time and associated costs. However, the higher number of endodontic treatments performed would have a financial impact.

In the Shadow of the 1919 Total Solar Eclipse: The Two British Expeditions and the Politics of Invisibility.

This paper addresses the legendary total solar eclipse of 29 May 1919. Two British teams confirmed the light bending prediction by Albert Einstein: Charles R. Davidson and Andrew C. C. Crommelin in Sobral, Brazil and Arthur S. Eddington and Edwin T. Cottingham on the African island of Príncipe, then part of the Portuguese empire. By jointly analyzing the two astronomical expeditions supported by written and visual sources, I show how, despite extensive scholarship on this famous historical episode and the historiographical emphasis on the plural dimensions of knowledge construction, many human and non-human actors have been kept in the shadow of the eclipse. I do so by focusing on what I call knowledge from the periphery together with knowledge from below, grounded literally on how localities (sites) affect choices and events, and growing outward to encompass a wide range of participants. I show how the geopolitical status of the two nations where the observational sites were located, and specifically Portugal's condition of colonial power, affected main decisions and events, while highlighting the active role of participants, ranging from experts from the peripheries and those involved in the travels to local elites and anonymous peoples, some of whom contributed to the observation of totality.

Purinergic signaling orchestrating neuron-glia communication.

This review discusses the evidence supporting a role for ATP signaling (operated by P2X and P2Y receptors) and adenosine signaling (mainly operated by A1 and A2A receptors) in the crosstalk between neurons, astrocytes, microglia and oligodendrocytes. An initial emphasis will be given to the cooperation between adenosine receptors to sharpen information salience encoding across synapses. The interplay between ATP and adenosine signaling in the communication between astrocytes and neurons will then be presented in context of the integrative properties of the astrocytic syncytium, allowing to implement heterosynaptic depression processes in neuronal networks. The process of microglia 'activation' and its control by astrocytes and neurons will then be analyzed under the perspective of an interplay between different P2 receptors and adenosine A2A receptors. In spite of these indications of a prominent role of purinergic signaling in the bidirectional communication between neurons and glia, its therapeutical exploitation still awaits obtaining an integrated view of the spatio-temporal action of ATP signaling and adenosine signaling, clearly distinguishing the involvement of both purinergic signaling systems in the regulation of physiological processes and in the control of pathogenic-like responses upon brain dysfunction or damage.

Quantitative ultrasound elastography and biometry of the bitch uterus in the early puerperium after vaginal delivery and caesarean section.

Elastography is an actual imaging method used to qualitatively and quantitatively evaluate the elastic properties of tissues. The aim of this study was to evaluate and compare uterine tissue biometry and stiffness during post-partum period in brachycephalic bitches (n = 12) after c-section (GCS; n = 8) or normal delivery (GNB; n = 4). These animals were evaluated daily by abdominal ultrasound from the day of delivery until the 10th day post-partum; measuring uterine diameter, myometrial and endometrial thickness (mm) and shear wave velocity (SWV; m/s), by B-mode and ARFI (acoustic radiation force impulse) elastography, respectively. Uterine diameter was higher (p = .012) in animals submitted to c-section (15.26 ± 4.73 mm) than in normal birth (12.53 ± 2.64 mm) during the first 7 days post-partum. Uterine thickness gradually involuted in both groups (p < .0001), the myometrium during the first 9 days (p = .005) and the endometrium during the first 6 days (p = .003). The myometrial and endometrial SWVs were similar between types of delivery (p = .7846 and .8273) and presented a gradual increase (p = .411; .0043, respectively), during the first 10 days post-partum. It was concluded that bitches with normal delivery had smaller uterine thickness and faster puerperal involution than submitted to c-section, while uterine tissue stiffness was similar between delivery types and increased gradually during post-partum.

Psychological pain and suicidal ideation in undergraduates: The role of pain avoidance.

In the present study, the first with a Western sample, we tested the relationship between three dimensions of psychological pain and suicidal ideation in a sample of 331 Portuguese college students. When controlling for the effect of depressive symptoms, the avoidance dimension, but not the cognitive and affective dimensions of psychological pain, made a significant contribution in predicting the three indicators of suicidal ideation. According to these results, psychotherapeutic action may focus either on enabling clients to tolerate mental pain or on providing more effective and adaptive strategies to cope with this pain.

The multidimensionality of suicide risk factors and criteria in a nonclinical population: Replication across two countries and two languages.

Across two countries and two languages, this research examined the multidimensional associations between suicidality (e.g., past ideation/attempts, communication of intent) and empirically important psychological risk factors (e.g., mental pain, perceived burdensomeness, thwarted belongingness). For samples of 228 Canadian and 331 Portuguese university undergraduates, four dimensions emerged in each sample with two of these, intrapersonal and interpersonal, demonstrating strong replicability across countries and languages. It was concluded that suicidality is a phenomenon that demonstrates some multidimensional similarities across cultures.

Adenosine A1 and A2A receptors differently control synaptic plasticity in the mouse dorsal and ventral hippocampus.

The hippocampus is a brain region involved in processing both memory and emotions, through a preferential involvement of the dorsal hippocampus (DH) and ventral hippocampus (VH), respectively. Adenosine A1 and A2A receptors (A1 R and A2A R) control both mood and memory, but it is not known if there is a different adenosine modulation of synaptic plasticity along the hippocampal axis. Using adult, C57BL/6 male mice, we show that both A1 R and A2A R were more abundant in DH compared with VH. However, recordings of field excitatory postsynaptic potentials at Schaffer collaterals-CA1 pyramidal synapses revealed that A1 R were equi-effective to inhibit basal excitatory synaptic transmission in DH and VH, but endogenous A1 R activation was more effective to depress the probability of release in VH. In contrast, the selective A2A R antagonist (SCH58261, 50 nM) controlled both long-term potentiation (induced by a high frequency stimulation protocol) and long-term depression (induced by a low frequency stimulation protocol) selectively in DH rather than VH, whereas the selective A1 R antagonist (DPCPX, 100 nM) revealed a similar tonic inhibition of long-term depression in DH and VH. These findings show a different control of synaptic plasticity by the adenosine modulation system in the dorsal and ventral poles of the hippocampus, which may underlie a different efficiency of the adenosine system to control mood and memory.

Acoustic radiation force impulse (ARFI) elastography to asses maternal and foetal structures in pregnant ewes.

The aim of this study was to evaluate the stiffness of foetal lungs, liver, kidneys and placentomes by acoustic radiation force impulse (ARFI) elastography in ovine and to correlate obtained findings with foetal development. Twenty-four ewes were included, and tissue stiffness of foetal lung, liver, kidney and placentome was measured beginning with 10th gestational week by qualitative-quantitative ARFI elastography. A total of 33 healthy lambs were born. Qualitative elastographic analysis permitted to classify maternal and foetal tissues elasticity in decreasing order as follows: placentome, kidney, liver and lung. Regarding quantitative ARFI elastography data, shear wave velocity (SWV) of foetal lung and liver varied. The lung SWV decreased gradually from the 16th to the 21st gestational week (R2  = 0.80; p < 0.001), while liver SWV increased gradually from the 14th to 21st gestational week (R2  = 0.80; p < 0.001). In contrast, the kidneys and placentomes SWV's remained constant through gestation (p = 0.076; and 0.34). ARFI elastography was shown feasible for evaluation of maternal and foetal tissues stiffness in the ovine model. It can be used to demonstrate pulmonary and hepatic stiffness modifications during foetal development.

Adenosine A2A receptors modulate the dopamine D2 receptor-mediated inhibition of synaptic transmission in the mouse prefrontal cortex.

Prefrontal cortex (PFC) circuits are modulated by dopamine acting on D1 - and D2 -like receptors, which are pharmacologically exploited to manage neuropsychiatric conditions. Adenosine A2A receptors (A2A R) also control PFC-related responses and A2A R antagonists are potential anti-psychotic drugs. As tight antagonistic A2A R-D2 R and synergistic A2A R-D1 R interactions occur in other brain regions, we now investigated the crosstalk between A2A R and D1 /D2 R controlling synaptic transmission between layers II/III and V in mouse PFC coronal slices. Dopamine decreased synaptic transmission, a presynaptic effect based on the parallel increase in paired-pulse responses. Dopamine inhibition was prevented by the D2 R-like antagonist sulpiride but not by the D1 R antagonist SCH23390 and was mimicked by the D2 R agonist sumanirole, but not by the agonists of either D4 R (A-412997) or D3 R (PD128907). Dopamine inhibition was prevented by the A2A R antagonist, SCH58261, and attenuated in A2A R knockout mice. Accordingly, triple-labelling immunocytochemistry experiments revealed the co-localization of A2A R and D2 R immunoreactivity in glutamatergic (vGluT1-positive) nerve terminals of the PFC. This reported positive A2A R-D2 R interaction controlling PFC synaptic transmission provides a mechanistic justification for the anti-psychotic potential of A2A R antagonists.

Caffeine alleviates progressive motor deficits in a transgenic mouse model of spinocerebellar ataxia.

OBJECTIVE: Machado-Joseph disease (MJD) is a neurodegenerative spinocerebellar ataxia (SCA) associated with an expanded polyglutamine tract within ataxin-3 for which there is currently no available therapy. We previously showed that caffeine, a nonselective adenosine receptor antagonist, delays the appearance of striatal damage resulting from expression of full-length mutant ataxin-3. Here we investigated the ability of caffeine to alleviate behavioral deficits and cerebellar neuropathology in transgenic mice with a severe ataxia resulting from expression of a truncated fragment of polyglutamine-expanded ataxin-3 in Purkinje cells. METHODS: Control and transgenic c57Bl6 mice expressing in the mouse cerebella a truncated form of human ataxin-3 with 69 glutamine repeats were allowed to freely drink water or caffeinated water (1g/L). Treatments began at 7 weeks of age, when motor and ataxic phenotype emerges in MJD mice, and lasted up to 20 weeks. Mice were tested in a panel of locomotor behavioral paradigms, namely rotarod, beam balance and walking, pole, and water maze cued-platform version tests, and then sacrificed for cerebellar histology. RESULTS: Caffeine consumption attenuated the progressive loss of general and fine-tuned motor function, balance, and grip strength, in parallel with preservation of cerebellar morphology through decreasing the loss of Purkinje neurons and the thinning of the molecular layer in different folia. Caffeine also rescued the putative striatal-dependent executive and cognitive deficiencies in MJD mice. INTERPRETATION: Our findings provide the first in vivo demonstration that caffeine intake alleviates behavioral disabilities in a severely impaired animal model of SCA. Ann Neurol 2017;81:407-418.

Unravelling Endogenous MicroRNA System Dysfunction as a New Pathophysiological Mechanism in Machado-Joseph Disease.

Machado-Joseph disease (MJD) is a genetic neurodegenerative disease caused by an expanded polyglutamine tract within the protein ataxin-3 (ATXN3). Despite current efforts, MJD's mechanism of pathogenesis remains unclear and no disease-modifying treatment is available. Therefore, in this study, we investigated (1) the role of the 3' UTR of ATXN3, a putative microRNA (miRNA) target, (2) whether miRNA biogenesis and machinery are dysfunctional in MJD, and (3) which specific miRNAs target ATXN3-3' UTR and whether they can alleviate MJD neuropathology in vivo. Our results demonstrate that endogenous miRNAs, by targeting sequences in the 3' UTR, robustly reduce ATXN3 expression and aggregation in vitro and neurodegeneration and neuroinflammation in vivo. Importantly, we found an abnormal MJD-associated downregulation of genes involved in miRNA biogenesis and silencing activity. Finally, we identified three miRNAs-mir-9, mir-181a, and mir-494-that interact with the ATXN3-3' UTR and whose expression is dysregulated in human MJD neurons and in other MJD cell and animal models. Furthermore, overexpression of these miRNAs in mice resulted in reduction of mutATXN3 levels, aggregate counts, and neuronal dysfunction. Altogether, these findings indicate that endogenous miRNAs and the 3' UTR of ATXN3 play a crucial role in MJD pathogenesis and provide a promising opportunity for MJD treatment.