Counting Synapses Using FIB/SEM Microscopy: A True Revolution for Ultrastructural Volume Reconstruction

The advent of transmission electron microscopy (TEM) in the 1950s represented a fundamental step in the study of neuronal circuits. The application of this technique soon led to the realization that the number of synapses changes during the course of normal life, as well as under certain pathological or experimental circumstances. Since then, one of the main goals in neurosciences has been to define simple and accurate methods to estimate the magnitude of these changes. Contrary to analysing single sections, TEM reconstructions are extremely time-consuming and difficult. Therefore, most quantitative studies use stereological methods to define the three-dimensional characteristics of synaptic junctions that are studied in two dimensions. Here, to count the exact number of synapses per unit of volume we have applied a new three-dimensional reconstruction method that involves the combination of focused ion beam milling and scanning electron microscopy (FIB/SEM). We show that the images obtained with FIB/SEM are similar to those obtained with TEM, but with the advantage that FIB/SEM permits serial reconstructions of large volumes of tissue to be generated rapidly and automatically. Furthermore, we compared the estimates of the number of synapses obtained with stereological methods with the values obtained by FIB/SEM reconstructions. We concluded that FIB/SEM not only provides the actual number of synapses per volume but it is also much easier and faster to use than other currently available TEM methods. More importantly, it also avoids most of the errors introduced by stereological methods and overcomes the difficulties associated with these techniques

Effects of Plyometric Jump Training in Sand or Rigid Surface on Jump-Related Biomechanical Variables and Physical Fitness in Female Volleyball Players

Background: This study aims to assess the effects of 8 weeks of plyometric jump training (PJT) conducted on sand or a rigid court surface on jump-related biomechanical variables and physical fitness in female indoor volleyball players. Methods: Seventeen participants were randomly divided into a sand surface group (SsG, n = 8) and rigid surface group (RsG, n = 9). Both groups completed equal indoor volleyball training routines. Participants were assessed pre and post the 8-week PJT for jump-related biomechanical variables (countermovement jump (CMJ) RSI; drop jump (DJ) reactive strength index (RSI); spike jump (SJ) height; CMJ height; CMJ rate of force development (RFD); CMJ velocity at take-off; DJ height and CMJ peak force), 20 m linear sprint time, t test for change-of-direction sprint (CODs) time, Wingate test peak power (PP), cardiorespiratory endurance, and leg-press one-repetition maximum (1RM). Results: A two-way mixed analysis of variance (group × time) revealed that there was a significant group × time interaction between DJ height (p = 0.035) and CMJ peak force (p = 0.032) in favour of RsG and SsG, respectively. A significant interaction was also observed for cardiorespiratory endurance (p = 0.01) and 1RM (p = 0.002), both favouring the SsG. No other group × time interaction was observed. Conclusions: The type of surface used during PJT induced specific adaptations in terms of jump-related biomechanical variables and physical fitness in female indoor volleyball players. Based on the individual needs of the athletes, practitioners may prescribe one type of surface preferentially over another to maximize the benefits derived from PJT

The Role of Pediatricians in Providing Greater-Quality Care for Children: An Ongoing Debate

Child healthcare frequently is regarded as a secondary objective in the development of public health policies and not as a central issue. Cost-containment policies instead of quality of healthcare frequently have inspired strategic decisions in public health investments for pediatric structures and workforce. The negative implications of this decisional approach have been shown by the substantial unpreparedness of virtually all European health systems during the coronavirus disease 2019 (COVID-19) pandemic,5 particularly in the area of primary care. The debate on the negative effects of cost-containment policies in the area of primary care in not new. In recent years, the role of pediatricians within the several European healthcare systems was variously reformed by governments, with an eye more to the budget than to the quality of care and its structural components, including staffing adequacy, training, and consistency as well as facility environment and size.1 As a result, the number of general pediatricians in Europe has decreased during the past 20 years.Data from the World Health Organization show that the average number of general pediatricians per 100 000 population in Europe declined 30.1% from 26.5% in 1998 to 18.5% in 2013.A further reduction in the pediatric workforce was reported in 2018 by the European Paediatric Association, the Union of National European Paediatric Societies and Associations (EPA-UNEPSA). During the past 20 years, due to different factors, including socioeconomic and political reasons, several European countries decided to reform their public health systems and the responsibility of pediatric healthcare delivery was moved from pediatricians to general and family practitioners. The aim of this commentary by the Spanish Primary Care Pediatrics Association (AEPap), in collaboration with the European Confederation of Primary Care Pediatricians and EPA-UNEPSA, is to discuss data supporting the essential role played in Europe by well-trained pediatricians vs other providers in delivering quality healthcare for children

Toward the Development of Efficient Electro-Fenton Reactors for Soil Washing Wastes through Microfluidic Cells

This work focuses on the coupling of three different approaches into the same reactor at the same time: microfluidic cells, anodic oxidation with diamond anodes, and an electro-Fenton process. To supply oxygen a jet aerator was used and a CB/PTFE Duocel Al foam cathode was installed to promote the formation of hydrogen peroxide. This novel concept is applied for the direct treatment (without the addition of salts or other reagents) of soil washing wastes obtained in the remediation of soil spiked with clopyralid. Results obtained pointed out that this approach can increase the efficiency of the process by folds as compared to traditional treatment technologies. The chemical analysis of the intermediates showed different reaction mechanisms: anodic oxidation, electro-Fenton, and a negligible contribution of coagulation. The coupled systems studied in this work present several advantages such as high treatment efficiency and short treatment time which indicate that the development of electrochemical reactors for diluted liquid wastes is progressing in an adequate direction.Este trabajo se centra en el acoplamiento de tres enfoques diferentes en el mismo reactor al mismo tiempo: celdas de microfluidos, oxidación anódica con ánodos de diamante y un proceso de electro-Fenton. Para suministrar oxígeno se utilizó un aireador de chorro y se instaló un cátodo de espuma CB / PTFE Duocel Al para promover la formación de peróxido de hidrógeno. Este novedoso concepto se aplica para el tratamiento directo (sin la adición de sales u otros reactivos) de los residuos del lavado de suelos obtenidos en la remediación de suelos enriquecidos con clopiralida. Los resultados obtenidos señalaron que este enfoque puede incrementar la eficiencia del proceso por pliegues en comparación con las tecnologías de tratamiento tradicionales. El análisis químico de los intermedios mostró diferentes mecanismos de reacción: oxidación anódica, electro-Fenton y una contribución insignificante de la coagulación

Understanding ozone generation in electrochemical cells at mild pHs

In this work, the production of ozone at mild pH conditions using a commercial electrochemical PEM cell CONDIAPURE Ⓡ is evaluated, at once a phenomenological model is proposed to understand the basis of the processes that occur inside the cell. At these pH conditions, the production of ozone can be explained from the oxidation of water, while the decomposition of ozone is found to be extremely important to explain the global experimental behavior observed. Not only is this decomposition a chemical but also an electrochemically assisted process which, in turn, can be related to the production of other oxidants in the cell which interact with ozone behaving as predators. The model formulated explains and satisfactorily reproduces the influence of the operation mode, the current intensity applied and presence and destruction of organics, with regression coefficients (r2) ranging from 0.88 to 0.99, helping to understand how the production of ozone should be promoted during electrochemical processes

Is ozone production able to explain the good performance of CabECO® technology in wastewater treatment?

This work intends to go a step further in the application of electrochemical technology to produce ozone, by evaluating the production of ozone with a commercial cell registered as CabECO®, which has been previously used to disinfect heavily polluted water in a successful manner, with a view to ascertaining whether this success may be related to the production of ozone. The CabECO® cell is a PEM electrolyzer; that is to say, one in which anode and cathode are connected through a PEM membrane. Since ozone production on the electrolyte is important and depends on the composition of the liquid, attention must be paid to the electrochemical production of scavengers. Dosing of oxygen as microbubbles has a very positive effect on the production of ozone, while temperature regulation is as well considerably important, as the increase in temperature generated by ohmic losses can lead to important decreases in the efficiency of ozone electrogeneration. Rates within the range 0.06–0.60 mg O3 min−1 are observed; this means that the maximum current efficiency reached is as high as 6.11% (considering oxygen as raw matter). These results are extremely promising as compared with other references in the literature, at once they also indicate there is room for further improvements and a long way to prevent the effect of scavengers and increase the efficiency of the process

Scale-up in PEM electro-ozonizers for the degradation of organics

This work focuses on the scale-up of electro-ozonizers by evaluating the production of ozone and the degradation of clopyralid synthetic wastes using three commercial PEM electrolyzers. The mechanical concept of the three cells is similar: a single compartment cell equipped with a MEA (consisting of a polymer exchange membrane and two pressed diamond coatings electrodes), powered with monopolar electric connection and where water flows on the surface of the electrodes, although the main electrolyte is the Nafion proton exchange membrane. However, their size and recommended operating conditions are not as similar, and their comparison becomes a good scaleup case study. The CabECO® cell consists of 2 MEAs with a total surface area of 24 cm2, a maximum operating current density of 2. 0A. The Mikrozom® cell consists of only one MEA with a net surface electrodic area of 112 mm2 and a maximum operation current density of 1.0 A. Finally, the CONDIAPURE® cell consists of a single MEA with a total surface area of 146 cm2 and a maximum operation current density of 10.0 A. The performance under mild and extreme operating conditions was compared and the results show that, although the cell concept is similar, the results obtained differ very significantly. The three PEM electrolyzers tested can produce ozone efficiently and mineralize completely clopyralid. The only intermediates measured come from the cathodic hydrodechlorination of clopyralid and oxidative intermediates were only detected at trace concentrations. CabECO® cell demonstrates an outstanding performance with very high current efficiencies in the production of ozone. However, the highest mineralization efficiencies are obtained with the Microzon®, which, although it is the PEM electrolyzer with the smallest active area, is the most efficient because can reach high ozone concentrations and achieve the best clopyralid mineralization. Efficiencies as high as 0.47 mg O3 Wh−1 can be obtained with this cell. Slightly lower values are reached by the CabECO® cell (0.38 mg O3 Wh−1). Enlarging electrode surface area does not seem to be a good strategy from the viewpoint of efficiency and it seems to promote side reactions that compete with ozone production and with the degradation of organics. This means that stacking rather than electrode enlarging should be the strategy more advisable for scaling up the electro-ozonation technology.Este trabajo se centra en la ampliación de los electro-ozonizadores mediante la evaluación de la producción de ozono y la degradación de los desechos sintéticos de clopiralida utilizando tres electrolizadores PEM comerciales. El concepto mecánico de las tres celdas es similar: una celda de un solo compartimiento equipada con un MEA (compuesto por una membrana de intercambio de polímero y dos electrodos de revestimiento de diamante prensado), alimentada con conexión eléctrica monopolar y donde el agua fluye en la superficie de los electrodos, aunque el electrolito principal es la membrana de intercambio de protones Nafion. Sin embargo, su tamaño y las condiciones operativas recomendadas no son tan similares, y su comparación se convierte en un buen caso de estudio de ampliación. La celda CabECO® consta de 2 MEA con una superficie total de 24 cm 2, una densidad de corriente operativa máxima de 2. 0A. La celda Mikrozom® consta de un solo MEA con un área superficial neta de electrodos de 112 mm 2 y una densidad de corriente máxima de operación de 1,0 A. Finalmente, la celda CONDIAPURE® consta de un solo MEA con un área superficial total de 146 cm 2y una densidad de corriente máxima de operación de 10.0 A. Se comparó el desempeño en condiciones de operación suaves y extremas y los resultados muestran que, aunque el concepto de celda es similar, los resultados obtenidos difieren muy significativamente. Los tres electrolizadores PEM probados pueden producir ozono de manera eficiente y mineralizar completamente la clopiralida. Los únicos intermedios medidos provienen de la hidrodecloración catódica de clopiralida y los intermedios oxidativos solo se detectaron en concentraciones mínimas. La celda CabECO® demuestra un desempeño sobresaliente con eficiencias de corriente muy altas en la producción de ozono. Sin embargo, las mayores eficiencias de mineralización se obtienen con el Microzon® que, si bien es el electrolizador PEM con menor área activa, es el más eficiente porque puede alcanzar altas concentraciones de ozono y lograr la mejor mineralización de clopiralida. Eficiencias de hasta 0,47 mg OCon esta celda se pueden obtener 3 Wh −1 . La celda CabECO® alcanza valores ligeramente inferiores (0,38 mg O 3 Wh −1 ). Ampliar el área de la superficie del electrodo no parece ser una buena estrategia desde el punto de vista de la eficiencia y parece promover reacciones secundarias que compiten con la producción de ozono y con la degradación de compuestos orgánicos. Esto significa que el apilamiento en lugar de la ampliación de electrodos debería ser la estrategia más recomendable para escalar la tecnología de electro-ozonización

RNF43 mutations predict response to anti-BRAF/EGFR combinatory therapies in BRAFV600E metastatic colorectal cancer

Colorectal cancer; Predictive markers; Tumour biomarkersCáncer colorrectal; Marcadores predictivos; Biomarcadores tumoralesCàncer colorrectal; Marcadors predictius; Biomarcadors tumoralsAnti-BRAF/EGFR therapy was recently approved for the treatment of metastatic BRAFV600E colorectal cancer (mCRCBRAF-V600E). However, a large fraction of patients do not respond, underscoring the need to identify molecular determinants of treatment response. Using whole-exome sequencing in a discovery cohort of patients with mCRCBRAF-V600E treated with anti-BRAF/EGFR therapy, we found that inactivating mutations in RNF43, a negative regulator of WNT, predict improved response rates and survival outcomes in patients with microsatellite-stable (MSS) tumors. Analysis of an independent validation cohort confirmed the relevance of RNF43 mutations to predicting clinical benefit (72.7% versus 30.8%; P = 0.03), as well as longer progression-free survival (hazard ratio (HR), 0.30; 95% confidence interval (CI), 0.12–0.75; P = 0.01) and overall survival (HR, 0.26; 95% CI, 0.10–0.71; P = 0.008), in patients with MSS-RNF43mutated versus MSS-RNF43wild-type tumors. Microsatellite-instable tumors invariably carried a wild-type-like RNF43 genotype encoding p.G659fs and presented an intermediate response profile. We found no association of RNF43 mutations with patient outcomes in a control cohort of patients with MSS-mCRCBRAF-V600E tumors not exposed to anti-BRAF targeted therapies. Overall, our findings suggest a cross-talk between the MAPK and WNT pathways that may modulate the antitumor activity of anti-BRAF/EGFR therapy and uncover predictive biomarkers to optimize the clinical management of these patients.VHIO would like to acknowledge the Cellex Foundation for providing research facilities and equipment, the FERO Foundation for their funding support, the Consorcio Centro de Investigación Biomédica en Red de Cáncer (CIBERONC, CB16/12/00259) from the Institute of Health Carlos III (ISCIII), Ministry of Science and Innovation, and the Department of Health (Generalitat de Catalunya, SLT008/18/00198 and SLT008/18/00205) for their support on this research. Authors acknowledge financial support from the State Agency for Research (Agencia Estatal de Investigación) (CEX2020-001024-S / AEI / 10.13039 /501100011033). This research is funded by the SCITRON program; Novartis funded the genomics characterization by WES of samples from 28 patients from the discovery cohort and had no influence on data analysis/interpretation or writing of the paper (3003145512 to R.A.T.). S.Li. is financially supported by a Chinese Scholarship Council PhD fellowship (201909370083 to S. Li). R.P.-L. is supported by a CRIS Foundation Talent Award (TALENT19-05), the FERO Foundation, the Instituto de Salud Carlos III-Investigación en Salud (PI18/01395 and PI21/01019 to R.P.-L.) and the Prostate Cancer Foundation (Young Investigator Award). This work was supported by the Miguel Servet-I Research Award from ISCIII of the Ministry of Economy (CP17/00199 to R.A.T.), the Olga Torres Foundation Award to emerging researchers (2601 to R.A.T.), the ISCIII-FEDER (PI17/00947 and PI20/00968 to E.E.), and the Fundación AECC (CLSEN19001ELEZ to E.E.) and Ministry of Science and Innovation (Europa Redes y Gestores, ECT2020-000827 to E.E.)

State of the art of audio- and video based solutions for AAL

Working Group 3. Audio- and Video-based AAL ApplicationsIt is a matter of fact that Europe is facing more and more crucial challenges regarding health and social care due to the demographic change and the current economic context. The recent COVID-19 pandemic has stressed this situation even further, thus highlighting the need for taking action. Active and Assisted Living (AAL) technologies come as a viable approach to help facing these challenges, thanks to the high potential they have in enabling remote care and support. Broadly speaking, AAL can be referred to as the use of innovative and advanced Information and Communication Technologies to create supportive, inclusive and empowering applications and environments that enable older, impaired or frail people to live independently and stay active longer in society. AAL capitalizes on the growing pervasiveness and effectiveness of sensing and computing facilities to supply the persons in need with smart assistance, by responding to their necessities of autonomy, independence, comfort, security and safety. The application scenarios addressed by AAL are complex, due to the inherent heterogeneity of the end-user population, their living arrangements, and their physical conditions or impairment. Despite aiming at diverse goals, AAL systems should share some common characteristics. They are designed to provide support in daily life in an invisible, unobtrusive and user-friendly manner. Moreover, they are conceived to be intelligent, to be able to learn and adapt to the requirements and requests of the assisted people, and to synchronise with their specific needs. Nevertheless, to ensure the uptake of AAL in society, potential users must be willing to use AAL applications and to integrate them in their daily environments and lives. In this respect, video- and audio-based AAL applications have several advantages, in terms of unobtrusiveness and information richness. Indeed, cameras and microphones are far less obtrusive with respect to the hindrance other wearable sensors may cause to one’s activities. In addition, a single camera placed in a room can record most of the activities performed in the room, thus replacing many other non-visual sensors. Currently, video-based applications are effective in recognising and monitoring the activities, the movements, and the overall conditions of the assisted individuals as well as to assess their vital parameters (e.g., heart rate, respiratory rate). Similarly, audio sensors have the potential to become one of the most important modalities for interaction with AAL systems, as they can have a large range of sensing, do not require physical presence at a particular location and are physically intangible. Moreover, relevant information about individuals’ activities and health status can derive from processing audio signals (e.g., speech recordings). Nevertheless, as the other side of the coin, cameras and microphones are often perceived as the most intrusive technologies from the viewpoint of the privacy of the monitored individuals. This is due to the richness of the information these technologies convey and the intimate setting where they may be deployed. Solutions able to ensure privacy preservation by context and by design, as well as to ensure high legal and ethical standards are in high demand. After the review of the current state of play and the discussion in GoodBrother, we may claim that the first solutions in this direction are starting to appear in the literature. A multidisciplinary 4 debate among experts and stakeholders is paving the way towards AAL ensuring ergonomics, usability, acceptance and privacy preservation. The DIANA, PAAL, and VisuAAL projects are examples of this fresh approach. This report provides the reader with a review of the most recent advances in audio- and video-based monitoring technologies for AAL. It has been drafted as a collective effort of WG3 to supply an introduction to AAL, its evolution over time and its main functional and technological underpinnings. In this respect, the report contributes to the field with the outline of a new generation of ethical-aware AAL technologies and a proposal for a novel comprehensive taxonomy of AAL systems and applications. Moreover, the report allows non-technical readers to gather an overview of the main components of an AAL system and how these function and interact with the end-users. The report illustrates the state of the art of the most successful AAL applications and functions based on audio and video data, namely (i) lifelogging and self-monitoring, (ii) remote monitoring of vital signs, (iii) emotional state recognition, (iv) food intake monitoring, activity and behaviour recognition, (v) activity and personal assistance, (vi) gesture recognition, (vii) fall detection and prevention, (viii) mobility assessment and frailty recognition, and (ix) cognitive and motor rehabilitation. For these application scenarios, the report illustrates the state of play in terms of scientific advances, available products and research project. The open challenges are also highlighted. The report ends with an overview of the challenges, the hindrances and the opportunities posed by the uptake in real world settings of AAL technologies. In this respect, the report illustrates the current procedural and technological approaches to cope with acceptability, usability and trust in the AAL technology, by surveying strategies and approaches to co-design, to privacy preservation in video and audio data, to transparency and explainability in data processing, and to data transmission and communication. User acceptance and ethical considerations are also debated. Finally, the potentials coming from the silver economy are overviewed.publishedVersio

High resolution human leukocyte antigen (HLA) class I and class II allele typing in Mexican mestizo women with sporadic breast cancer: case-control study

<p>Abstract</p> <p>Background</p> <p>The development of breast cancer is multifactorial. Hormonal, environmental factors and genetic predisposition, among others, could interact in the presentation of breast carcinoma. Human leukocyte antigen (HLA) alleles play an important role in immunity (cellular immunity) and may be important genetic traits. HLAAllele-specific interaction has not been well established. Recently, several studies had been conducted in order to do so, but the results are controversial and in some instances contradictory.</p> <p>Methods</p> <p>We designed a case-control study to quantify the association of HLA class I and II genes and breast cancer. HLA typing was performed by high resolution sequence-specific oligotyping after DNA amplification (PCR-SSOP) of 100 breast cancer Mexican mestizo patients and 99 matched healthy controls.</p> <p>Results</p> <p>HLA-A frequencies that we were able to observe that there was no difference between both groups from the statistical viewpoint. HLA-B*1501 was found three times more common in the case group (OR, 3.714; <it>p </it>= 0.031). HLA-Cw is not a marker neither for risk, nor protection for the disease, because we did not find significant statistical differences between the two groups. DRB1*1301, which is expressed in seven cases and in only one control, observing an risk increase of up to seven times and DRB1*1602, which behaves similarly in being present solely in the cases (OR, 16.701; 95% CI, 0.947 – 294.670). DQ*0301-allele expression, which is much more common in the control group and could be protective for the presentation of the disease (OR, 0.078; 95% CI, 0.027–0.223, <it>p </it>= 0.00001).</p> <p>Conclusion</p> <p>Our results reveal the role of the MHC genes in the pathophysiology of breast cancer, suggesting that in the development of breast cancer exists a disorder of immune regulation. The triggering factor seems to be restricted to certain ethnic groups and certain geographical regions since the relevant MHC alleles are highly diverse. This is the first study in Mexican population where high resolutions HLA typing has been performed in order to try to establish an association with malignancy.</p