Circulating extracellular vesicle proteins and microRNA profiles in subcortical and cortical-subcortical ischaemic stroke

In order to investigate the role of circulating extracellular vesicles (EVs), proteins, and microRNAs as damage and repair markers in ischaemic stroke depending on its topography, subcor-tical (SC), and cortical-subcortical (CSC) involvement, we quantified the total amount of EVs using an enzyme-linked immunosorbent assay technique and analysed their global protein content using proteomics. We also employed a polymerase chain reaction to evaluate the circulating microRNA profile. The study included 81 patients with ischaemic stroke (26 SC and 55 CSC) and 22 healthy controls (HCs). No differences were found in circulating EV levels between the SC, CSC, and HC groups. We detected the specific expression of C1QA and Casp14 in the EVs of patients with CSC ischaemic stroke and the specific expression of ANXA2 in the EVs of patients with SC involvement. Patients with CSC ischaemic stroke showed a lower expression of miR-15a, miR-424, miR-100, and miR-339 compared with those with SC ischaemic stroke, and the levels of miR-339, miR-100, miR-199a, miR-369a, miR-424, and miR-15a were lower than those of the HCs. Circulating EV proteins and microRNAs from patients with CSC ischaemic stroke could be considered markers of neurite outgrowth, neurogenesis, inflammation process, and atherosclerosis. On the other hand, EV proteins and microRNAs from patients with SC ischaemic stroke might be markers of an anti-inflammatory process and blood–brain barrier disruption reduction.This work was sponsored by a grant from Miguel Servet (CP15/00069; CPII20/00002 to María Gutiérrez-Fernández), Miguel Servet (CP20/00024 to Laura Otero-Ortega), a predoctoral fellowship (FI17/00188 to Mari Carmen Gómez-de Frutos; FI18/00026 to Fernando Laso-García), a Sara Borrell postdoctoral fellowship (CD19/00033 to María Pérez-Mato), a Río Hortega (CM20/00047 to Elisa Alonso-López) and the INVICTUS PLUS network grant (RD16/0019/0005) from the Carlos III Health Institute Health Care Research Fund and was co-funded by the European Regional Development Fund (ERDF)

Quorum sensing network in clinical strains of A. baumannii: AidA is a new quorum quenching enzyme

18 p.-4 fig.-6 tab. López, María et al.Acinetobacter baumannii is an important pathogen that causes nosocomial infections generally associated with high mortality and morbidity in Intensive Care Units (ICUs). Currently, little is known about the Quorum Sensing (QS)/Quorum Quenching (QQ) systems of this pathogen. We analyzed these mechanisms in seven clinical isolates of A. baumannii. Microarray analysis of one of these clinical isolates, Ab1 (A. baumannii ST-2_clon_2010), previously cultured in the presence of 3-oxo-C12-HSL (a QS signalling molecule) revealed a putative QQ enzyme (α/ß hydrolase gene, AidA). This QQ enzyme was present in all non-motile clinical isolates (67% of which were isolated from the respiratory tract) cultured in nutrient depleted LB medium. Interestingly, this gene was not located in the genome of the only motile clinical strain growing in this medium (A. baumannii strain Ab421_GEIH-2010 [Ab7], isolated from a blood sample). The AidA protein expressed in E. coli showed QQ activity. Finally, we observed downregulation of the AidA protein (QQ system attenuation) in the presence of H2O2 (ROS stress). In conclusion, most of the A. baumannii clinical strains were not surface motile (84%) and were of respiratory origin (67%). Only the pilT gene was involved in surface motility and related to the QS system. Finally, a new QQ enzyme (α/ß hydrolase gene, AidA protein) was detected in these strains.This study was funded by grant PI13/02390 awarded to MT within the State Plan for R+D+I 2013-2016 (National Plan for Scientific Research, Technological Development and Innovation 2008-2011) and co-financed by the ISCIII-Deputy General Directorate of evaluation and Promotion of Research - European Regional Development Fund "A way of Making Europe" and Instituto de Salud Carlos III FEDER, Spanish Network for the Research in Infectious Diseases (REIPI RD12/0015).Peer reviewe

Similarities and differences in extracellular vesicle profiles between ischaemic stroke and myocardial infarction

Extracellular vesicles (EVs) are involved in intercellular signalling through the transfer of molecules during physiological and pathological conditions, such as ischaemic disease. EVs might therefore play a role in ischaemic stroke (IS) and myocardial infarction (MI). In the present study, we analysed the similarities and differences in the content of circulating EVs in patients with IS and MI. This prospective observational study enrolled 140 participants (81 patients with IS, 37 with MI and 22 healthy controls [HCs]). We analysed the protein and microRNA content from EVs using proteomics and reverse transcription quantitative real-time polymerase chain reaction and compared it between the groups. In the patients with IS and MI, we identified 14 common proteins. When comparing IS and MI, we found differences in the protein profiles (apolipoprotein B, alpha-2-macroglobulin, fibronectin). We also found lower levels of miR-340 and miR-424 and higher levels of miR-29b in the patients with IS and MI compared with the HCs. Lastly, we found higher miR-340 levels in IS than in MI. In conclusion, proteomic and miRNA analyses suggest a relationship between circulating EV content and the patient’s disease state. Although IS and MI affect different organs (brain and heart) with distinct histological characteristics, certain EV proteins and miRNAs appear to participate in both diseases, while others are present only in patients with ISThis work was sponsored by a grant from Miguel Servet (CP15/00069 to María Gutiérrez- Fernández), a predoctoral fellowship (FI17/00188 to Mari Carmen Gómez-de Frutos;FI18/00026 to Fernando Laso-García), a Sara Borrell postdoctoral fellowship (CD19/00033 to María Pérez-Mato), the INVICTUS PLUS network grant (RD16/0019/0005) from the Carlos III Health Institute Health Care Research Fund and was co-funded by the European Regional Development Fund (ERDF) and Juan de la Cierva postdoctoral fellowship (IJCI-2017-33505 to Laura Otero-Ortega, Spanish State Research Agency) and the Spanish Ministry of Science and Innovatio

Coping with intimate partner violence and the COVID-19 lockdown: The perspectives of service professionals in Spain

Socioeconomic crisis and humanitarian disasters can cause increased stress for women who experience inter-partner violence (IPV). This study analyzed the impact of the COVID-19 lockdown on this important issue, their related health and social services and working conditions from the perspectives of professionals in different sectors. Forty-three semi-structured interviews were carried out with 47 professionals (44 women and 3 men) from 40 different entities (September 2020-April 2021). This content analysis suggests that the pandemic and its associated prevention measures have had a negative impact on women exposed to IPV and their children, which affected their social wellbeing. Professionals described burnout, difficult and slow administrative processes, and problems with coordination and access to information. These negative impacts were mitigated, in part, by the work of professionals, but this suggests that a series of key strategies are needed to improve the response capacity of the service sector to IPV in situations of crisis. These improvements are related to the availability of human and material resources; an efficient coordination network between the professionals from different sectors; existence of informal support networks in the community; protocols/procedures and prior training for better implementation; and greater flexibility and accessibility of basic services that benefit women who experience IPV.This study was financed through the project “Gender violence and social and health responses during the COVID-19 crisis” by the Fondo Supera Covid-19 CRUE-Santander for the period 2020-2021 (Ref. FSCovid19-03). It was also co-supported by the CIBER of Epidemiology and Public Health of Spain for its aid to the Gender-based Violence and Youth Research Program.S

White matter repair after extracellular vesicles administration in an experimental animal model of subcortical stroke

Mesenchymal stem cells have previously been shown to mediate brain repair after stroke; they secrete 50-100 nm complexes called extracellular vesicles (EVs), which could be responsible for provoking neurovascular repair and functional recovery. EVs have been observed by electron microscopy and NanoSight, and they contain associated proteins such as CD81 and Alix. This purified, homogeneous population of EVs was administered intravenously after subcortical stroke in rats. To evaluate the EVs effects, we studied the biodistribution, proteomics analysis, functional evaluation, lesion size, fiber tract integrity, axonal sprouting and white matter repair markers. We found that a single administration of EVs improved functional recovery, fiber tract integrity, axonal sprouting and white matter repair markers in an experimental animal model of subcortical stroke. EVs were found in the animals' brain and peripheral organs after euthanasia. White matter integrity was in part restored by EVs administration mediated by molecular repair factors implicated in axonal sprouting, tract connectivity, remyelination and oligodendrogenesis. These findings are associated with improved functional recovery. This novel role for EVs presents a new perspective in the development of biologics for brain repair.This study has been partially supported by grants from PS15/01318 and INVICTUS (RD12/0014) (Spanish Neurovascular Network), Miguel Servet (CP15/00069 to María Gutiérrez-Fernández) and a Sara Borrell postdoctoral fellowship (CD12/00706, to Laura Otero-Ortega) from Research Institute Carlos III, Ministry of Science and Innovation of Spain and European Regional Development Fun

Whole-Exome sequencing reveals recurrent but heterogeneous mutational profiles in sporadic who grade 1 meningiomas

© 2021 González-Tablas, Prieto, Arandia, Jara-Acevedo, Otero, Pascual, Ruíz, Álvarez-Twose, García-Montero, Orfao and Tabernero.Human WHO grade 1 meningiomas are generally considered benign tumors; despite this, they account for ≈50% of all recurrent meningiomas. Currently, limited data exist about the mutational profiles of grade 1 meningiomas and patient outcome. We investigated the genetic variants present in 32 WHO grade 1 meningiomas using whole exome sequencing, and correlated gene mutational profiles with tumor cytogenetics and patient outcome. Overall, WHO grade 1 meningiomas harbored numerous and heterogeneous genetic variants, which most frequently affected the NF2 (47%) gene and to a less extent the PNMA6A (22%), TIGD1 (16%), SMO (13%), PTEN (13%), CREG2 (9%), EEF1A1 (6%), POLR2A (6%), ARID1B (3%), and FAIM3 (3%) genes. Notably, non-synonymous genetic variants of SMO and POLR2A were restricted to diploid meningiomas, whereas NF2 mutations were only found among tumors that showed -22/22q─ (with or without a complex karyotype). Based on NF2 mutations and tumor cytogenetics, four genetic profiles were defined with an impact on patient recurrence-free survival (RFS). These included (1) two good-prognosis tumor subgroups—diploid meningiomas (n=9) and isolated -22/22q─ associated with NF2 mutation (n=7)—with RFS rates at 10 y of 100%; and (2) two subgroups of poor-prognosis meningiomas—isolated -22/22q─ without NF2 mutation (n=3) and tumors with complex karyotypes (n=11)—with a RFS rate at 10 y of 48% (p=0.003). Our results point out the existence of recurrent but heterogeneous mutational profiles in WHO grade 1 meningiomas which have an impact on patient outcome.This work was supported by grants: IBY 17/00002 from IBSAL (Salamanca, Spain), GRS2132/A2020 from Junta de Castilla y León (Spain), and CB16/12/00400 from CIBER-ONC and FONDOS FEDER (Instituto de Salud Carlos III, Ministerio de Economía y Competitividad, Madrid, Spain)

The role of ultrasound as a diagnostic and therapeutic tool in experimental animal models of stroke: A review

Ultrasound is a noninvasive technique that provides real-time imaging with excellent resolution, and several studies demonstrated the potential of ultrasound in acute ischemic stroke monitoring. However, only a few studies were performed using animal models, of which many showed ultrasound to be a safe and effective tool also in therapeutic applications. The full potential of ultrasound application in experimental stroke is yet to be explored to further determine the limitations of this technique and to ensure the accuracy of translational research. This review covers the current status of ultrasound applied to monitoring and treatment in experimental animal models of stroke and examines the safety, limitations, and future perspectives.This research was funded by the Carlos III Health Institute Health Care Research Fund grant number FIS PI16/01052 and cofunded by the European Regional Development Fund (ERDF)–Miguel Servet (CP15/00069, CPII20/00002 to María Gutiérrez–Fernández; CP20/00024 to Laura Otero–Ortega) and predoctoral fellowship (FI17/00188 to Mari Carmen Gómez–de Frutos, FI18/00026 to Fernando Laso–García) and the INVICTUS PLUS Spanish Network (RD16/0019/0005) of the Carlos III Health Institute (ISCIII)

Isolasi Bakteri Pendegradasi Senyawa Persisten Organic Pollutants Asal Tanah Inceptisol Karawang

Penggunaan pestisida yang tidak terkontrol akan mengganggu agroekosistem pertanian dan mencemari lingkungan. Jumlah pestisida yang beredar di Indonesia tahun 2006 terdaftar sebanyak 1336 formulasi, 2008 sebanyak 1702 formulasi, 2010 sebanyak 2048 formulasi, 2011 sebanyak 2247 formulasi. Di dalam tanah, karbon aktif peranan sebagai shelter atau rumah untuk mikroorganisme. Pori-pori kecil pada karbon aktif digunakan sebagai tempat tinggal bakteri, sedangkan pori besar dan retakan (cracks) digunakan sebagai tempat berkumpul. Penelitian dilakukan di Laboratorium Mikrobiologi Pusat Penelitian Biologi LIPI Cibinong Mei 2011 s/d Agustus 2011.Tujuan penelitian adalah menyeleksi bakteri dalam tanah yang berpotensi mendegradasi residu insektisida yang bersifat persistent organic poluttants (POPs). Isolasi dan karakterisasi mikroba pendegradasi POPs dilakukan dengan tiga tahapan, yaitu (1) Isolasi dan identifikasi mikroba pendegradasi POPs, (2) Uji karakteristik pertumbuhan isolat pada berbagai jenis POPs, dan (3) Penetapan residu insektisida POPs hasil kultur. Hasil terdapat 7 (tujuh) isolat yang mampu mendegradasi senyawa POPs, Lima isolat bersifat gram positif (BOB1, BOB2, BOB3, BOB4, BOB5) efektif untuk mendegradasi POPs berbahan aktif: lindan, heptaklor, DDT, dan dieldrin, sedangkan dua isolat bersifat gram negatif (BOB6 dan BOB7) efektif untuk mendegradasi POPs berbahan aktif aldrin

Pharmacokinetics of Intravitreal Anti-VEGF Drugs in Age-Related Macular Degeneration

Intravitreal administration of anti-vascular endothelial growth factor (VEGF) antibodies has become the standard treatment for Age-Related Macular Degeneration; however, the knowledge of their pharmacokinetics is limited. A comprehensive review of the preclinical and clinical pharmacokinetic data that were obtained in different studies with intravitreal bevacizumab, ranibizumab, and aflibercept has been conducted. Moreover, the factors that can influence the vitreous pharmacokinetics of these drugs, as well as the methods that were used in the studies for analytical determination, have been exposed. These anti-VEGF drugs present different charge and molecular weights, which play an important role in vitreous distribution and elimination. The pharmacokinetic parameters that were collected differ depending on the species that were involved in the studies and on physiological and pathological conditions, such as vitrectomy and lensectomy. Knowledge of the intravitreal pharmacokinetics of the anti-VEGF drugs that were used in clinical practice is of vital importance.This work was partially supported by the ISCIII (PI17/00940, RETICS Oftared, RD16/0008/0003 and RD12/0034/0017) cofunded by FEDER and by the Spanish Ministry of Science, Innovation and Universities (RTI2018-099597-B-100)S

Exosomes promote restoration after an experimental animal model of intracerebral hemorrhage

Exosomes are gaining importance because they show great promise in therapeutic applications for several diseases. Particularly in stroke, exosomes derived from mesenchymal stem cell (MSC) therapy work as paracrine effectors responsible for promoting neurovascular remodeling and functional recovery. Adult male rats were subjected to intracerebral hemorrhage (ICH) by intrastriatal injection of collagenase type IV; 24 h after surgery, MSC-derived exosomes were administered through the tail vein. The rats were euthanized at 7 or 28 days after treatment. Functional evaluation, lesion size, fiber tract integrity, axonal sprouting and white matter repair markers, biodistribution of exosomes and secretome proteomics were analyzed. DiI-labeled exosomes were found in the brains of the ICH-treated group and in the liver, lung and spleen. Animals receiving treatment with exosomes showed significantly better results in terms of functional recovery, lesion size, fiber tract integrity, axonal sprouting and white matter repair markers compared with the control group 28 days after stroke. Proteomics analysis of the exosomes identified more than 2000 proteins that could be implicated in brain repair function. In conclusion, white matter integrity was partly restored by exosome administration mediated by molecular repair factors. Exosomes as a treatment could be a heterogeneous process by nature and presents many factors that can influence brain plasticity in an adaptable and versatile manner.The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This study has been partially supported by grants from PS15/01318 and INVICTUS (RD12/0014) (Spanish Neurovascular Network), Miguel Servet (CP15/00069 to Marıa Gutierrez-Fernandez) and a Sara Borrell postdoctoral fellowship (CD12/00706, to Laura Otero-Ortega) from the Research Institute Carlos III, the Spanish Ministry of Science and Innovation and the European Regional Development Fund. This study was also co-funded within the Roche Farma project. The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.S