Hybrid Fluorescent Mass-Tag Nanotrackers as Universal Reagents for Long-Term Live-Cell Barcoding.

Barcoding and pooling cells for processing as a composite sample are critical to minimize technical variability in multiplex technologies. Fluorescent cell barcoding has been established as a standard method for multiplexing in flow cytometry analysis. In parallel, mass-tag barcoding is routinely used to label cells for mass cytometry. Barcode reagents currently used label intracellular proteins in fixed and permeabilized cells and, therefore, are not suitable for studies with live cells in long-term culture prior to analysis. In this study, we report the development of fluorescent palladium-based hybrid-tag nanotrackers to barcode live cells for flow and mass cytometry dual-modal readout. We describe the preparation, physicochemical characterization, efficiency of cell internalization, and durability of these nanotrackers in live cells cultured over time. In addition, we demonstrate their compatibility with standardized cytometry reagents and protocols. Finally, we validated these nanotrackers for drug response assays during a long-term coculture experiment with two barcoded cell lines. This method represents a new and widely applicable advance for fluorescent and mass-tag barcoding that is independent of protein expression levels and can be used to label cells before long-term drug studies.

Active psychodynamic psychotherapy: moments of high receptiveness in working with emotions.

Moments of High Receptiveness (MoHR) offer support to active dynamic psychotherapy.

Development and Implementation of a Hybrid Wireless Sensor Network of Low Power and Long Range for Urban Environments.

The urban population, worldwide, is growing exponentially and with it the demand for information on pollution levels, vehicle traffic, or available parking, giving rise to citizens connected to their environment. This article presents an experimental long range (LoRa) and low power consumption network, with a combination of static and mobile wireless sensors (hybrid architecture) to tune and validate concentrator placement, to obtain a large coverage in an urban environment. A mobile node has been used, carrying a gateway and various sensors. The Activation By Personalization (ABP) mode has been used, justified for urban applications requiring multicasting. This allows to compare the coverage of each static gateway, being able to make practical decisions about its location. With this methodology, it has been possible to provide service to the city of Malaga, through a single concentrator node. The information acquired is synchronized in an external database, to monitor the data in real time, being able to geolocate the dataframes through web mapping services. This work presents the development and implementation of a hybrid wireless sensor network of long range and low power, configured and tuned to achieve efficient performance in a mid-size city, and tested in experiments in a real urban environment.

The Internet of Cooperative Agents Architecture (X-IoCA) for Robots, Hybrid Sensor Networks, and MEC Centers in Complex Environments: A Search and Rescue Case Study.

Cloud robotics and advanced communications can foster a step-change in cooperative robots and hybrid wireless sensor networks (H-WSN) for demanding environments (e.g., disaster response, mining, demolition, and nuclear sites) by enabling the timely sharing of data and computational resources between robot and human teams. However, the operational complexity of such multi-agent systems requires defining effective architectures, coping with implementation details, and testing in realistic deployments. This article proposes X-IoCA, an Internet of robotic things (IoRT) and communication architecture consisting of a hybrid and heterogeneous network of wireless transceivers (H2WTN), based on LoRa and BLE technologies, and a robot operating system (ROS) network. The IoRT is connected to a feedback information system (FIS) distributed among multi-access edge computing (MEC) centers. Furthermore, we present SAR-IoCA, an implementation of the architecture for search and rescue (SAR) integrated into a 5G network. The FIS for this application consists of an SAR-FIS (including a path planner for UGVs considering risks detected by a LoRa H-WSN) and an ROS-FIS (for real-time monitoring and processing of information published throughout the ROS network). Moreover, we discuss lessons learned from using SAR-IoCA in a realistic exercise where three UGVs, a UAV, and responders collaborated to rescue victims from a tunnel accessible through rough terrain.

Integration of a Mobile Node into a Hybrid Wireless Sensor Network for Urban Environments.

Robots, or in general, intelligent vehicles, require large amounts of data to adapt their behavior to the environment and achieve their goals. When their missions take place in large areas, using additional information to that gathered by the onboard sensors frequently offers a more efficient solution of the problem. The emergence of Cyber-Physical Systems and Cloud computing allows this approach, but integration of sensory information, and its effective availability for the robots or vehicles is challenging. This paper addresses the development and implementation of a modular mobile node of a Wireless Sensor Network (WSN), designed to be mounted onboard vehicles, and capable of using different sensors according to mission needs. The mobile node is integrated with an existing static network, transforming it into a Hybrid Wireless Sensor Network (H-WSN), and adding flexibility and range to it. The integration is achieved without the need for multi-hop routing. A database holds the data acquired by both mobile and static nodes, allowing access in real-time to the gathered information. A Human⁻Machine Interface (HMI) presents this information to users. Finally, the system is tested in real urban scenarios in a use-case of measurement of gas levels.

Analysis of 3D Scan Measurement Distribution with Application to a Multi-Beam Lidar on a Rotating Platform.

Multi-beam lidar (MBL) rangefinders are becoming increasingly compact, light, and accessible 3D sensors, but they offer limited vertical resolution and field of view. The addition of a degree-of-freedom to build a rotating multi-beam lidar (RMBL) has the potential to become a common solution for affordable rapid full-3D high resolution scans. However, the overlapping of multiple-beams caused by rotation yields scanning patterns that are more complex than in rotating single beam lidar (RSBL). In this paper, we propose a simulation-based methodology to analyze 3D scanning patterns which is applied to investigate the scan measurement distribution produced by the RMBL configuration. With this purpose, novel contributions include: (i) the adaption of a recent spherical reformulation of Ripley's K function to assess 3D sensor data distribution on a hollow sphere simulation; (ii) a comparison, both qualitative and quantitative, between scan patterns produced by an ideal RMBL based on a Velodyne VLP-16 (Puck) and those of other 3D scan alternatives (i.e., rotating 2D lidar and MBL); and (iii) a new RMBL implementation consisting of a portable tilting platform for VLP-16 scanners, which is presented as a case study for measurement distribution analysis as well as for the discussion of actual scans from representative environments. Results indicate that despite the particular sampling patterns given by a RMBL, its homogeneity even improves that of an equivalent RSBL.

Voxel-Based Neighborhood for Spatial Shape Pattern Classification of Lidar Point Clouds with Supervised Learning.

Improving the effectiveness of spatial shape features classification from 3D lidar data is very relevant because it is largely used as a fundamental step towards higher level scene understanding challenges of autonomous vehicles and terrestrial robots. In this sense, computing neighborhood for points in dense scans becomes a costly process for both training and classification. This paper proposes a new general framework for implementing and comparing different supervised learning classifiers with a simple voxel-based neighborhood computation where points in each non-overlapping voxel in a regular grid are assigned to the same class by considering features within a support region defined by the voxel itself. The contribution provides offline training and online classification procedures as well as five alternative feature vector definitions based on principal component analysis for scatter, tubular and planar shapes. Moreover, the feasibility of this approach is evaluated by implementing a neural network (NN) method previously proposed by the authors as well as three other supervised learning classifiers found in scene processing methods: support vector machines (SVM), Gaussian processes (GP), and Gaussian mixture models (GMM). A comparative performance analysis is presented using real point clouds from both natural and urban environments and two different 3D rangefinders (a tilting Hokuyo UTM-30LX and a Riegl). Classification performance metrics and processing time measurements confirm the benefits of the NN classifier and the feasibility of voxel-based neighborhood.

Hirschsprung disease with debut in adult age as acute intestinal obstruction: case report.

Hirschsprung's disease is characterized by absence of ganglion cells in submucosal and myenteric plexus of distal bowel. Most cases become manifest during the neonatal period, but in rare instances, this disease is initially diagnosed in adult age. It usually presents as severe constipation with colonic dilatation proximal to the aganglionic segment. The treatment is surgical, removing the aganglionic segment and restoring continuity of digestive tract. The disease rarely presents as an acute intestinal obstruction. We report a case not previously diagnosed, which presented as a massive colonic dilatation with a maximum diameter of 44 cm, with imminent risk of drilling that forced to perform an emergency surgery. We include a review of existing literature.

Assessment of heavy metal pollution in Córdoba (Spain) by biomonitoring foraging honeybee.

Due to features that make them outstanding environmental bioindicator, colonies of Apis mellifera are being used to study environmental pollution. The primary objective of this research was to use honeybee colonies to identify heavy metals and determine their utility for environmental management. Five stations each with two A. mellifera hives were strategically located in urban, industrial, agricultural and forested areas within the municipality of Córdoba (Spain), and foraging bees were collected from April to December in 2007, 2009 and 2010 to analyse spatial and temporal variation in Pb, Cr, Ni and Cd pollution. Metal concentrations, in milligram per kilogram of honeybee, were determined by inductively coupled plasma-atomic emission spectrometry and graphite furnace atomic absorption spectrophotometry. Significant differences in concentrations were found among the various locations and periods. The highest number of values exceeding the upper reference thresholds proposed for this study (Pb, 0.7 mg/kg; Cr, 0.12 mg/kg; Ni, 0.3 mg/kg; and Cd, 0.1 mg/kg) was observed for Pb and Cr (6.25% respectively), station S4 (13.22%), year 2007 (20.83%) and in months of May and July (11.90% each). Regarding the Cd, which was analysed only in 2010, the highest number of values exceeding the upper reference thresholds was 40%. Biomonitoring with colonies of A. mellifera could contribute to improved surveillance and control systems for atmospheric pollution by integrating qualitative and quantitative assessments, thus facilitating prevention and readiness in the event of environmental crises.

Active Change in Psychodynamic Therapy: Moments of High Receptiveness.

This article presents the concept of "moments of high receptiveness" (MoHR or "Momentos de Alta Receptividad"), which is derived from the concept of "experiential coupling" ("Acoplamiento de Experiencias") proposed by Bleichmar (2001). Experiential coupling recently received empirical support by the work of Schiller and colleagues (2010). We will also show the conceptual placing of moments of high receptiveness with respect to the developments of Stern and colleagues (Stern and et al., 1998; Stern, 2004). In order to achieve both objectives, we focus on various clinical vignettes stressing the differences in repercussions of the technique. We describe use of stimuli for active evocation, explain how to identify moments of high receptiveness, and review ways to take advantage of these moments. Lastly, to minimize the risk of iatrogenic symptoms, we examine the role of therapists and some features of the therapeutic process when using this technique.

[Development of the surgical approach to acute cholecystitis in an emergency surgical unit]. / Evolución del abordaje quirúrgico de la colecistitis aguda en una unidad de cirugía de urgencias.

OBJECTIVE: The application of the laparoscopic technique in acute cholecystitis is still subject to controversy. The aim of this study is to asses the applicability, safety, benefits and complications of the laparoscopic approach in patients with acute cholecystitis, as well as the development of this technique in the emergency surgery department of a tertiary hospital, compared to laparotomy. MATERIAL AND METHOD: The study consisted of 354 patients with acute cholecystitis syndromes operated either by open or laparoscopic surgery, during the years 2006 to 2009. RESULTS: The laparoscopic method was used in 253 patients, and 101 by the open route, with the slight majority being male (57.67%) and with a mean age of 62.83 years. The number of laparoscopic cholecystectomies increased from 60% in 2006, to 79% in 2009. The mean hospital stay (including those with and without complications) was shorter using the laparoscopic approach, compared to open surgery (showing a difference of approximately 6 days). The postoperative complications in laparoscopy during the four years studied decreased from 21.42 to 11.3%. The local and general complications were significantly associated with time since the start of the acute symptoms and the surgery, as well as the histopathological state of the gall bladder. CONCLUSIONS: The laparoscopic approach continues to play an increasing role in the treatment of this disease, becoming the main surgical option in our hospital.

The effect of being housed with adult female flock-mates on the age at puberty of autumn-born ewe-lambs.

Twenty-five ewe-lambs born in November were used to measure the effect of the presence of cyclic or non-cyclic adult ewes on the age at first fertile estrus. Lambs were weaned at 45 days of age and kept in a single flock until 3 Mar (4 months old), when they were assigned to one of the following three groups: Group CE (n = 8), housed with six cyclic adult ewes; Group NCE (n = 8), housed with three non-cyclic ovariectomized adult ewes, and Group ISO (n = 9), housed isolated from adult ewes. On 2nd May (6 months old), two rams were introduced into each group, until 31st Oct (12 months old), when rams were removed. Adult ewes in group CE were separated from rams and ewe-lambs by a metal fence, allowing visual, olfactory, and nose-to-nose contact between animals. Average earliest fertile estrus was significantly (P < 0.01) earlier in NCE ewe-lambs (8th Aug) than it was in ISO (9th Sep) and CE (17th Sep) ewe-lambs, they lambed about 35 d earlier (1st Jan) than did the other two groups (2nd Feb and 10th Feb, respectively) (P < 0.01), and at an earlier age (14 mo, 15.2 mo, and 15.3 mo, respectively; P < 0.01). In conclusion, the presence of non-ovulatory adult ewes advanced puberty in autumn-born ewe-lambs of a Mediterranean genotype. The factor(s) that delayed age at puberty in isolated females and those housed with estrous adult ewes remains to be elucidated.

Polymer film as starch azure container for the easy diastase activity determination in honey.

A new original application for a polyacrylic film based on the monomers 2-(dimethylamino)ethyl methacrylate (NNDA), 2-hydroxyethyl acrylate (2HEA) and methylmethacrylate (MMA) as a starch azure container has been set up for a simple determination of honey diastase activity. The proposed method is based on the correlation of reducing sugars generated during the enzymatic process with the Schade reference assay. The polyacrylic film is charged with starch azure acting as a container for this substance; thus, the starch does not interfere in the measurement of reducing sugars, so that the diastase activity is easily calculated. The method has been contrasted with Schade method, showing good correlation and differences under 0.4% between methods in some honey samples. The polyacrylic film has great potential for the routine honey diastase activity assessment in small laboratories, dramatically reducing analysis time and cost.

Effectiveness of aprepitant in post-acute COVID19 syndrome.

This is the first case of a patient taking aprepitant for a post-acute COVID-19 syndrome. This case may encourage researchers to look for the evidence for the efficacy and safety of a neurokinin 1 receptor antagonist in this frequent syndrome.

An effective polymeric nanocarrier that allows for active targeting and selective drug delivery in cell coculture systems.

In this manuscript, we report the development of a versatile, robust, and stable targeting nanocarrier for active delivery. This nanocarrier is based on bifunctionalized polymeric nanoparticles conjugated to a monoclonal antibody that allows for active targeting of either (i) a fluorophore for tracking or (ii) a drug for monitoring specific cell responses. This nanodevice can efficiently discriminate between cells in coculture based on the expression levels of cell surface receptors. As a proof of concept, we have demonstrated efficient delivery using a broadly established cell surface receptor as the target, the epidermal growth factor receptor (EGFR), which is overexpressed in several types of cancers. Additionally, a second validation of this nanodevice was successfully carried out using another cell surface receptor as the target, the cluster of differentiation 147 (CD147). Our results suggest that this versatile nanocarrier can be expanded to other cell receptors and bioactive cargoes, offering remarkable discrimination efficiency between cells with different expression levels of a specific marker. This work supports the ability of nanoplatforms to boost and improve the progress towards personalized medicine.

Role of the Non-Canonical RNAi Pathway in the Antifungal Resistance and Virulence of Mucorales.

Mucorales are the causal agents for the lethal disease known as mucormycosis. Mortality rates of mucormycosis can reach up to 90%, due to the mucoralean antifungal drug resistance and the lack of effective therapies. A concerning urgency among the medical and scientific community claims to find targets for the development of new treatments. Here, we reviewed different studies describing the role and machinery of a novel non-canonical RNAi pathway (NCRIP) only conserved in Mucorales. Its non-canonical features are the independence of Dicer and Argonaute proteins. Conversely, NCRIP relies on RNA-dependent RNA Polymerases (RdRP) and an atypical ribonuclease III (RNase III). NCRIP regulates the expression of mRNAs by degrading them in a specific manner. Its mechanism binds dsRNA but only cuts ssRNA. NCRIP exhibits a diversity of functional roles. It represses the epimutational pathway and the lack of NCRIP increases the generation of drug resistant strains. NCRIP also regulates the control of retrotransposons expression, playing an essential role in genome stability. Finally, NCRIP regulates the response during phagocytosis, affecting the multifactorial process of virulence. These critical NCRIP roles in virulence and antifungal drug resistance, along with its exclusive presence in Mucorales, mark this pathway as a promising target to fight against mucormycosis.

Assessing heavy metal pollution by biomonitoring honeybee nectar in Córdoba (Spain).

Nectar of honeybee colonies has been used in order to identify heavy metals and establish the benefit of this type of studies as a tool for environmental management. For these goals, samples of nectar were obtained from Apis mellifera hives placed in the city of Córdoba (Spain) and its surroundings. Five stations (each with two hives) were selected and samples were collected from May to July of 2007, 2009 and 2010. Concentrations of Pb, Cr, Ni and Cd in nectar were determined by graphite furnace atomic absorption spectrophotometry. Substantial spatial and temporal differences were detected and compared with the values found in bee bodies in a previously published study based on samples obtained simultaneously with those presented in this work. Upper reference thresholds established for this investigation were surpassed frequently by the measures obtained, being Cr (21.43% of samples), stations S3 (22.22%) and S4 (11.12%) year 2009 (22.22%) and the month of July (23.68%) the metal, the locations and the periods that exceeded more times these references. Regarding the Cd, which was studied only in 2010, 33.33% of the nectar samples exceeded the upper reference thresholds. Comparing the biomonitoring of bee bodies and nectar, some coincidences were found, although they showed different results for highest worrisome values of metal, station and year. This suggests that both methods can give complementary information in the surveillance systems of atmospheric pollution.

Characterization and Therapeutic Effect of a pH Stimuli Responsive Polymeric Nanoformulation for Controlled Drug Release.

Despite the large number of polymeric nanodelivery systems that have been recently developed, there is still room for improvement in terms of therapeutic efficiency. Most reported nanodevices for controlled release are based on drug encapsulation, which can lead to undesired drug leakage with a consequent reduction in efficacy and an increase in systemic toxicity. Herein, we present a strategy for covalent drug conjugation to the nanodevice to overcome this drawback. In particular, we characterize and evaluate an effective therapeutic polymeric PEGylated nanosystem for controlled pH-sensitive drug release on a breast cancer (MDA-MB-231) and two lung cancer (A549 and H520) cell lines. A significant reduction in the required drug dose to reach its half maximal inhibitory concentration (IC50 value) was achieved by conjugation of the drug to the nanoparticles, which leads to an improvement in the therapeutic index by increasing the efficiency. The genotoxic effect of this nanodevice in cancer cells was confirmed by nucleus histone H2AX specific immunostaining. In summary, we successfully characterized and validated a pH responsive therapeutic polymeric nanodevice in vitro for controlled anticancer drug release.

Genes, Pathways, and Mechanisms Involved in the Virulence of Mucorales.

The order Mucorales is a group of ancient fungi with limited tools for gene manipulation. The main consequence of this manipulation unwillingness is the limited knowledge about its biology compared to other fungal groups. However, the emerging of mucormycosis, a fungal infection caused by Mucorales, is attracting the medical spotlight in recent years because the treatments available are not efficient in reducing the high mortality associated with this disease. The result of this renewed interest in Mucorales and mucormycosis is an extraordinarily productive effort to unveil their secrets during the last decade. In this review, we describe the most compelling advances related to the genetic study of virulence factors, pathways, and molecular mechanisms developed in these years. The use of a few genetic study models has allowed the characterization of virulence factors in Mucorales that were previously described in other pathogens, such as the uptake iron systems, the mechanisms of dimorphism, and azole resistances. More importantly, recent studies are identifying new genes and mechanisms controlling the pathogenic potential of Mucorales and their interactions with the host, offering new alternatives to develop specific strategies against mucormycosis.

Peroxiredoxin 6 Down-Regulation Induces Metabolic Remodeling and Cell Cycle Arrest in HepG2 Cells.

Peroxiredoxin 6 (Prdx6) is the only member of 1-Cys subfamily of peroxiredoxins in human cells. It is the only Prdx acting on phospholipid hydroperoxides possessing two additional sites with phospholipase A2 (PLA2) and lysophosphatidylcholine-acyl transferase (LPCAT) activities. There are contrasting reports on the roles and mechanisms of multifunctional Prdx6 in several pathologies and on its sensitivity to, and influence on, the redox environment. We have down-regulated Prdx6 with specific siRNA in hepatoblastoma HepG2 cells to study its role in cell proliferation, redox homeostasis, and metabolic programming. Cell proliferation and cell number decreased while cell volume increased; import of glucose and nucleotide biosynthesis also diminished while polyamines, phospholipids, and most glycolipids increased. A proteomic quantitative analysis suggested changes in membrane arrangement and vesicle trafficking as well as redox changes in enzymes of carbon and glutathione metabolism, pentose-phosphate pathway, citrate cycle, fatty acid metabolism, biosynthesis of aminoacids, and Glycolysis/Gluconeogenesis. Specific redox changes in Hexokinase-2 (HK2), Prdx6, intracellular chloride ion channel-1 (CLIC1), PEP-carboxykinase-2 (PCK2), and 3-phosphoglycerate dehydrogenase (PHGDH) are compatible with the metabolic remodeling toward a predominant gluconeogenic flow from aminoacids with diversion at 3-phospohglycerate toward serine and other biosynthetic pathways thereon and with cell cycle arrest at G1/S transition.