Cabotegravir Exposure of Zebrafish (Danio rerio) Embryos Impacts on Neurodevelopment and Behavior

As most new medications, Cabotegravir (CAB) was recently approved as an antiretroviral treatment of HIV infection without in-depth safety information on in utero exposure. Although no developmental toxicity in rats and rabbits was reported, recent studies demonstrated that CAB decreases pluripotency of human embryonic stem cells. CAB exposure effects during development were assessed in zebrafish embryos by the Fish Embryo Toxicity test after exposure at subtherapeutic concentrations up to 25x the human C-max. Larvae behavior was assessed by the light-dark locomotion test. The expression of factors involved in neurogenesis was evaluated by whole-mount in situ hybridization. CAB did not cause gross morphological defects at low doses, although pericardial edema, uninflated swim bladder, decreased heartbeats, growth delay, and decreased hatching rate were observed at the highest concentrations. Decreased locomotion was observed even at the subtherapeutic dose, suggesting alterations of nervous system integrity. This hypothesis was supported by the observation of decreased expression of crucial factors involved in early neuronal differentiation in diencephalic and telencephalic dopaminergic areas, midbrain/hindbrain boundary, and craniofacial ganglia. These findings support CAB effects on neurogenesis in zebrafish embryos and suggest long-term follow-up of exposed infants to provide data on drug safety during pregnancy

Tracheostomy is associated with increased survival in Multiple System Atrophy patients with stridor

Stridor treatment in multiple system atrophy (MSA) mainly comprises tracheostomy or continuous positive airway pressure (CPAP), but guidelines for the use of these treatments are lacking. The aim of the study was to evaluate the predictive value of stridor treatment in an MSA cohort

Comparison of Efavirenz and Doravirine Developmental Toxicity in an Embryo Animal Model

: In the past, one of the most widely used non-nucleoside reverse transcriptase inhibitors (NNRTI) in first-line antiretroviral therapy (ART) of HIV infection was efavirenz (EFV), which is already used as a cost-effective treatment in developing countries due to its efficacy, tolerability, and availability. However, EFV also demonstrates several adverse effects, like hepatotoxicity, altered lipid profile, neuropsychological symptoms, and behavioral effects in children after in utero exposure. In 2018, another NNRTI, doravirine (DOR), was approved due to its similar efficacy but better safety profile. Preclinical safety studies demonstrated that DOR is not genotoxic and exhibits no developmental toxicity or effects on fertility in rats. Zebrafish (Danio rerio) embryos have been widely accepted as a vertebrate model for pharmacological and developmental studies. We used zebrafish embryos as an in vivo model to investigate the developmental toxicity of DOR compared to EFV. After exposure of the embryos to the drugs from the gastrula stage up to different developmental stages (30 embryos for each arm, in three independent experiments), we assessed their survival, morphology, hatching rate, apoptosis in the developing head, locomotion behavior, vasculature development, and neutral lipid distribution. Overall, DOR showed a better safety profile than EFV in our model. Therapeutic and supra-therapeutic doses of DOR induced very low mortality [survival rates: 92, 90, 88, 88, and 81% at 1, 5, 10, 25, and 50 μM, respectively, at 24 h post fecundation (hpf), and 88, 85, 88, 89, and 75% at the same doses, respectively, at 48 hpf] and mild morphological alterations compared to EFV exposure also in the sub-therapeutic ranges (survival rates: 80, 77, 69, 63, and 44% at 1, 5, 10, 25, and 50 μM, respectively, at 24 hpf and 72, 70, 63, 52, and 0% at the same doses, respectively, at 48 hpf). Further, DOR only slightly affected the hatching rate at supra-therapeutic doses (97, 98, 96, 87, and 83% at 1, 5, 10, 25, and 50 μM, respectively, at 72 hpf), while EFV already strongly reduced hatching at sub-therapeutic doses (83, 49, 11, 0, and 0% at 1, 5, 10, 25, and 50 μM, respectively, at the same time endpoint). Both DOR at therapeutic doses and most severely EFV at sub-therapeutic doses enhanced apoptosis in the developing head during crucial phases of embryo neurodevelopment and perturbed the locomotor behavior. Furthermore, EFV strongly affected angiogenesis and disturbed neutral lipid homeostasis even at sub-therapeutic doses compared to DOR at therapeutic concentrations. Our findings in zebrafish embryos add further data confirming the higher safety of DOR with respect to EFV regarding embryo development, neurogenesis, angiogenesis, and lipid metabolism. Further studies are needed to explore the molecular mechanisms underlying the better pharmacological safety profile of DOR, and further human studies are required to confirm these results in the zebrafish animal model

Modulation of the Muscle Activity During Sleep in Cervical Dystonia

Introduction: Impaired sleep has been reported as an important nonmotor feature in dystonia, but so far, self-reported complaints have never been compared with nocturnal video-polysomnographic (PSG) recording, which is the gold standard to assess sleep-related disorders. Methods: Twenty patients with idiopathic isolated cervical dystonia and 22 healthy controls (HC) underwent extensive clinical investigations, neurological examination, and questionnaire screening for excessive daytime sleepiness and sleep-related disorders. A full-night video PSG was performed in both patients and HC. An ad hoc montage, adding electromyographic leads over the muscle affected with dystonia, was used. Results: When compared to controls, patients showed significantly increased pathological values on the scale assessing self-reported complaints of impaired nocturnal sleep. Higher scores of impaired nocturnal sleep did not correlate with any clinical descriptors but for a weak correlation with higher scores on the scale for depression. On video-PSG, patients had significantly affected sleep architecture (with decreased sleep efficiency and increased sleep latency). Activity over cervical muscles disappears during all the sleep stages, reaching significantly decreased values when compared to controls both in nonrapid eye movements and rapid eye movements sleep. Conclusions: Patients with cervical dystonia reported poor sleep quality and showed impaired sleep architecture. These features however cannot be related to the persistence of muscle activity over the cervical muscles, which disappears in all the sleep stages, reaching significantly decreased values when compared to HC

Индекс стабилизации Фабри (FASTEX): инновационный инструмент для оценки клинической стабилизации при болезни Фабри

На сегодняшний день предложены 2 системы количественной оценки бремени гликогеноза с дефицитом α-галактозидазы: индекс оценки степени тяжести Майнца (MSSI) и система балльной оценки тяжести болезни Фабри (DS3). Сделана попытка разработать динамическую математическую модель FASTEX (от англ. FAbry STabilization indEX, индекс стабилизации Фабри) для оценки клинической стабильности состояния. Мультидисциплинарная группа экспертов по болезни Фабри впервые предложила новую шкалу оценки тяжести заболевания по предварительной оценке (от англ. raw score, RS), основанную на 3 доменах (домен нервной системы (боль, цереброваскулярные события), почечный домен (протеинурия, скорость клубочковой фильтрации), сердечный домен (параметры эхокардиографии, электрокардиографии и степень сердечной недостаточности по классификации Нью-Йоркской кардиологической ассоциации)) с небольшим числом пунктов в каждом из них и оценкой клинической стабильности во времени. RS протестирована на 28 пациентах (15 мужчин и 13 женщин) с классической формой болезни Фабри. Получена сильная корреляционная связь предложенной оценки RS и взвешенной оценки (от англ. weighted score, WS) с DS3 и MSSI (r2 = 0,914; 0,949; 0,910 и 0,938 соответственно). Для уточнения RS была рассчитана WS, выражаемая в процентах. WS была основана на относительной клинической значимости каждого пункта в пределах домена, при этом группа экспертов согласовывала присвоение разного веса клинического вреда конкретной системе органов. Для определения динамики тяжести заболевания RS была повторно определена через 1 год. Группа экспертов согласилась с пороговым ограничением в 20 % от исходного уровня в качестве клинической WS для определения клинической стабильности. Модель FASTEX показала хорошую корреляцию с клинической оценкой и клиническим изменением на протяжении времени у всех пациентов. 

The X-Ray Outburst of the Galactic Center Magnetar over Six Years of Chandra Observations

The magnetar SGR J1745−2900, discovered at a distance of parsecs from the Milky Way central black hole, Sagittarius A*, represents the closest pulsar to a supermassive black hole ever detected. Furthermore, its intriguing radio emission has been used to study the environment of the black hole, as well as to derive a precise position and proper motion for this object. The discovery of SGR J1745−2900 has led to interesting debates about the number, age, and nature of pulsars expected in the Galactic center region. In this work, we present extensive X-ray monitoring of the outburst of SGR J1745−2900 using the Chandra X-ray Observatory, the only instrument with the spatial resolution to distinguish the magnetar from the supermassive black hole (2"4 angular distance). It was monitored from its outburst onset in 2013 April until 2019 August, collecting more than 50 Chandra observations for a total of more than 2.3 Ms of data. Soon after the outburst onset, the magnetar emission settled onto a purely thermal emission state that cooled from a temperature of about 0.9–0.6 keV over 6 yr. The pulsar timing properties showed at least two changes in the period derivative, increasing by a factor of about 4 during the outburst decay. We find that the long-term properties of this outburst challenge current models for the magnetar outbursts.N.R., D.V., and A.B. are supported by the H2020 ERC Consolidator Grant “MAGNESIA” under grant agreement No. 817661 (PI: Rea). N.R., F.C.Z., D.V., A.B., and D.F.T. also acknowledge support from grants SGR2017-1383 and PGC2018-095512-BI00. F.C.Z. is supported by a Juan de la Cierva fellowship. A.P. acknowledges financial support from grants ASI/INAF I/037/12/0, ASI/INAF 2017-14-H.0 (PI: Belloni) and from INAF grant “Sostegno alla ricerca scientifica main streams dell’INAF,” Presidential Decree 43/2018 (PI: Belloni). D.H. acknowledges support from the Natural Sciences and Engineering Research Council of Canada (NSERC) Discovery Grant, the Fonds de recherche du Québec–Nature et Technologies (FRQNT) Nouveaux Chercheurs program, and the Canadian Institute for Advanced Research (CIFAR). G.L.I., S.M., and R.T. have been partially supported by PRIN-MIUR 2017. J.A.P. acknowledges support by the Generalitat Valenciana (PROMETEO/2019/071) and by Agencia Estatal de Investigación (PGC2018-095984-B-I00). G.P. is supported by the H2020 ERC Consolidator Grant “Hot Milk” under grant agreement No. 865637. L.S. acknowledges financial contributions from ASI-INAF agreements 2017-14-H.O and I/037/12/0 and from “iPeska” research grant (PI: Andrea Possenti) funded under the INAF call PRIN-SKA/CTA (resolution 70/2016). We acknowledge support from the PHAROS COST Action (CA16214)

Hyperactive Akt1 Signaling Increases Tumor Progression and DNA Repair in Embryonal Rhabdomyosarcoma RD Line and Confers Susceptibility to Glycolysis and Mevalonate Pathway Inhibitors

In pediatric rhabdomyosarcoma (RMS), elevated Akt signaling is associated with increased malignancy. Here, we report that expression of a constitutively active, myristoylated form of Akt1 (myrAkt1) in human RMS RD cells led to hyperactivation of the mammalian target of rapamycin (mTOR)/70-kDa ribosomal protein S6 kinase (p70S6K) pathway, resulting in the loss of both MyoD and myogenic capacity, and an increase of Ki67 expression due to high cell mitosis. MyrAkt1 signaling increased migratory and invasive cell traits, as detected by wound healing, zymography, and xenograft zebrafish assays, and promoted repair of DNA damage after radiotherapy and doxorubicin treatments, as revealed by nuclear detection of phosphorylated H2A histone family member X (γH2AX) through activation of DNA-dependent protein kinase (DNA-PK). Treatment with synthetic inhibitors of phosphatidylinositol-3-kinase (PI3K) and Akt was sufficient to completely revert the aggressive cell phenotype, while the mTOR inhibitor rapamycin failed to block cell dissemination. Furthermore, we found that pronounced Akt1 signaling increased the susceptibility to cell apoptosis after treatments with 2-deoxy-D-glucose (2-DG) and lovastatin, enzymatic inhibitors of hexokinase, and 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGCR), especially in combination with radiotherapy and doxorubicin. In conclusion, these data suggest that restriction of glucose metabolism and the mevalonate pathway, in combination with standard therapy, may increase therapy success in RMS tumors characterized by a dysregulated Akt signaling

Clinical Features and Pathophysiology of Disorders of Arousal in Adults: A Window Into the Sleeping Brain

Introduction: Disorders of Arousal (DoA) are NREM parasomnias that have been typically regarded as self-limited childhood manifestations. It is now clear that DoA can persist in adults, often presenting with distinctive characteristics. So far, few studies have described the clinical course and characteristics of DoA in adulthood, therefore a large part of their semiology is ignored. The aim of this study is to describe the clinical manifestations of DoA in an adult population and to provide a pathophysiological interpretation of their features.Methods: We screened our database for all 1,600 adult (≥15 years) patients with sleep-related motor behaviors between 1995 and 2016. We identified 45 patients with typical DoA episodes, of whom a complete history, neurological examination and diagnostic video-polysomnography (VPSG) were available. All patients provided a detailed description of their episodes (with particular regards to semiology, frequency, and association with stressful life events) in different life periods. VPSG recordings were reviewed and DoA episodes were identified and assigned to three different categories according to their complexity.Results: Our population was composed of 45 adult patients ranging between 15 and 76 years. Sleepwalking was reported by 86% of patients, possibly associated with complex interactions with the environment and violent behaviors in 53% of cases; distressing mental contents were reported by 64%. Recall of the episodes was reported in 77% of patients. Non-restorative sleep was reported in 46% of patients. Stress was a potential episode trigger in 80% of patients. VPSG recordings documented 334 DoA episodes. According to our classification of motor patterns, 282 episodes (84%) were Simple Arousal Movements (SAMs), 34 (10%) Rapid Arousal Movements (RAMs) and 18 (5%) Complex Arousal Movements (CAMs).Discussion: Our study confirms that DoA in adulthood present with distinctive characteristics, such as non-restorative sleep, violence and complex, or bizarre behaviors. Alternative classifications of DoA based on motor patterns could be useful to characterize DoA episodes in adults, as different motor patterns often coexist in the same individual and minor episodes are more common but generally underreported by patients. Prospective studies are needed for a definitive characterization of DoA in adulthood throughout the life course

The multi-outburst activity of the magnetar in Westerlund I

After two major outbursts in 2006 and 2011, on 2017 May 16 the magnetar CXOU J164710.2−455216, hosted within the massive star cluster Westerlund I, emitted a short (∼20 ms) burst, which marked the onset of a new active phase. We started a long-term monitoring campaign with Swift (45 observations), Chandra (five observations), and NuSTAR (four observations) from the activation until 2018 April. During the campaign, Swift Burst Alert Telescope (BAT) registered the occurrence of multiple bursts, accompanied by two other enhancements of the X-ray persistent flux. The long time span covered by our observations allowed us to study the spectral and the timing evolution of the source. After ∼11 months since the 2017 May outburst onset, the observed flux was ∼15 times higher than its historical minimum level and a factor of ∼3 higher than the level reached after the 2006 outburst. This suggests that the crust has not fully relaxed to the quiescent level, or that the source quiescent level has changed following the multiple outburst activities in the past 10 yr or so. This is another case of multiple outbursts from the same source on a yearly time-scale, a somehow recently discovered behaviour in magnetars.AB, NR, and PE are supported by an NWO Vidi Grant (PI: Rea). NR is also supported by grants AYA2015-71042-P and SGR 2014-1073. PE acknowledges funding in the framework of the project ‘Understanding the X-ray Variable and Transient Sky’ (ULTraS), ASI-INAF contract no. 2017-14-H.0. JAP acknowledges support by the Spanish MINECO/FEDER grant AYA2015-66899-C2-2-P, and the grant of Generalitat Valenciana PROMETEOII-2014-069. FCZ is supported by grants AYA2015-71042-P and SGR 2014-1073. DG acknowledges the financial support of the UnivEarthS Labex program at Sorbonne Paris Citeé (ANR-10-LABX-0023 and ANR-11-IDEX-0005-02). We thank the referee for his comments and the COST Action PHAROS (CA16214) for partial support

All-sky Medium Energy Gamma-ray Observatory: Exploring the Extreme Multimessenger Universe

The All-sky Medium Energy Gamma-ray Observatory (AMEGO) is a probe class mission concept that will provide essential contributions to multimessenger astrophysics in the late 2020s and beyond. AMEGO combines high sensitivity in the 200 keV to 10 GeV energy range with a wide field of view, good spectral resolution, and polarization sensitivity. Therefore, AMEGO is key in the study of multimessenger astrophysical objects that have unique signatures in the gamma-ray regime, such as neutron star mergers, supernovae, and flaring active galactic nuclei. The order-of-magnitude improvement compared to previous MeV missions also enables discoveries of a wide range of phenomena whose energy output peaks in the relatively unexplored medium-energy gamma-ray band