Multi Omics Applications in Biological Systems.

Traditional methodologies often fall short in addressing the complexity of biological systems. In this regard, system biology omics have brought invaluable tools for conducting comprehensive analysis. Current sequencing capabilities have revolutionized genetics and genomics studies, as well as the characterization of transcriptional profiling and dynamics of several species and sample types. Biological systems experience complex biochemical processes involving thousands of molecules. These processes occur at different levels that can be studied using mass spectrometry-based (MS-based) analysis, enabling high-throughput proteomics, glycoproteomics, glycomics, metabolomics, and lipidomics analysis. Here, we present the most up-to-date techniques utilized in the completion of omics analysis. Additionally, we include some interesting examples of the applicability of multi omics to a variety of biological systems.

Aminomethylphosphonic Acid (AMPA), a Glyphosate Metabolite, Decreases Plasma Cholinesterase Activity in Rats.

Glyphosate, a widely used herbicide, is linked to a plethora of deleterious effects in both clinical and preclinical studies. Nevertheless, the effects of its main metabolite, aminomethylphosphonic acid (AMPA), whose half-life in soil is even longer than that of glyphosate, have been little explored. On this basis, as a first approach, in this work, we report that intraperitoneal (i.p.) administration of AMPA or glyphosate (at 10, 56, and 100 mg/kg) decreased, to a similar extent, plasma cholinesterase (ChE) activity in acutely exposed rats. Moreover, we designed an experimental protocol to analyze and compare the effects of AMPA and glyphosate on human plasma ChE activity; this protocol consisted of adding these compounds to human plasma to subsequently test the effects of this plasma on the contraction to acetylcholine (ACh) in the frog rectus abdominis muscle (an indirect estimate of ChE activity). Accordingly, this muscular contraction to ACh was evaluated before and after pre-incubation of ACh with (i) plasma alone, (ii) plasma with AMPA, and (iii) plasma with glyphosate. Our results indicate that AMPA, like glyphosate, decreased ChE activity in the plasma of rats (when given i.p.) and humans (when added in vitro), suggesting that both xenobiotics may exert similar toxicological effects.

Both acute glyphosate and the aminomethylphosphonic acid intoxication decreased the acetylcholinesterase activity in rat hippocampus, prefrontal cortex and gastrocnemius muscle.

It has been reported that glyphosate, one of the most common herbicides used in agriculture, impairs locomotion and cognition. Glyphosate has a variable half-life in soil up to biotic and/or abiotic factors transform the molecule in metabolites such as the aminomethylphosphonic acid (AMPA) that has a longer half-life. In this study, female Sprague Dawley rats were acutely exposed to different doses of glyphosate or AMPA (i.e. 10, 56 or 100 mg/kg) and, subsequently, the acetylcholinesterase (AChE) activity was measured in the hippocampus, prefrontal cortex (PFC) and the gastrocnemius muscle. Both glyphosate and AMPA produced a similar decrease in the AChE activity in all the tissues tested. These results suggest that interference with normal cholinergic neurotransmission may be one of the mechanisms involved in glyphosate-induced motor alterations in rats. Moreover, our results highlight the biological importance of AMPA as a molecule with anticholinesterase action in brain and skeletal muscle. To our knowledge, this is the first report showing in vivo that AMPA, the major metabolite of glyphosate, behaves as an organophosphate.

Neurotoxicity of glyphosate: Focus on molecular mechanisms probably associated with alterations in cognition and behavior.

In recent decades, glyphosate and glyphosate-based herbicides (GBH) have been extensively used in agriculture all over the world. Initially, they were considered safe, but rising evidence suggests that these molecules reach the central nervous system producing metabolic, functional, and permanent alterations that impact cognition and behavior. This theoretical and non-systematic review involved searching, integrating, and analyzing preclinical evidence regarding the effects of acute, sub-chronic, and chronic exposure to glyphosate and GBH on cognition, behavior, neural activity, and development in adult and juvenile rodents following perinatal exposition. In addition, this review gathers the mechanisms underlying the neurotoxicity of glyphosate mediating cognitive and behavioral alterations. Furthermore, clinical evidence of the effects of exposition to GBH on human health and its possible link with several neurological disorders was revised.

Metabolomic Changes in Rat Serum after Chronic Exposure to Glyphosate-Based Herbicide.

Glyphosate-based herbicides (GBHs) have gained extensive popularity in recent decades. For many years, glyphosate has been regarded as harmless or minimally toxic to mammals due to the absence of its primary target, the shikimic acid pathway in humans. Nonetheless, mounting evidence suggests that glyphosate may cause adverse health effects in humans via other mechanisms. In this study, we described the metabolomic changes in the serum of experimental rats exposed to chronic GBH using the highly sensitive LC-MS/MS technique. We investigated the possible relationship between chronic exposure to GBH and neurological disorders. Our findings suggest that chronic exposure to GBH can alter spatial learning memory and the expression of some important metabolites that are linked to neurophysiological disorders in young rats, with the female rats showing higher susceptibility compared to the males. This indicates that female rats are more likely to show early symptoms of the disorder on exposure to chronic GBH compared to male rats. We observed that four important metabolites (paraxanthine, epinephrine, L-(+)-arginine, and D-arginine) showed significant changes and involvement in neurological changes as suggested by ingenuity pathway analysis. In conclusion, our results indicate that chronic exposure to GBH can increase the risk of developing neurological disorders.

Obtaining polyphenolic extracts from pomegranate peel (Punica granatum) to evaluate the bactericide and antioxidant activity.

Pomegranate (Punica granatum) contains secondary metabolites with antioxidant and bactericide activity; however, the study of the peel in the endemic varieties of Mexico has not been deepened. The polyphenols extraction of peel pomegranate endemic to the state of Michoacan, Mexico could be used in the formlulation of healthy food due contains antioxidant compounds or could be used like drugs due contains antibactericide compunds.In this work 3 varieties of pomegranate were analyzed; Wonderful, Apaseo and Tecozautla harvested in 2017 and 2018, carrying out a physicochemical characterization to establish the ripening, application of an experimental design of response surface for drying the peel and extracting polyphenols using two solvents (acetone and ethanol) by the Soxhlet method. As a result, the pomegranates were in the correct ripening, in the drying an optimal point of operation was found without affecting the metabolites (36 h at 55 °C) and in the extraction, the bactericide and antioxidant activity was evaluated observing that in the ketone extracts the best results were obtained in the Apaseo variety being; ABTS•+ technique of 150.78 ET mM/g, DPPH• 109.8 ET mM/g and 11.82 EAG mg/g in dry extract. For the bactericide activity measured by inhibition halos in S. aureus and E. coli it was had; 20.03 mm and 14.05 mm respectively for the Apaseo variety, which is why it is convenient to extract polyphenols under this method in peel of Mexican pomegranate varieties.

Involvement of the PRL-PAK1 Pathway in Cancer Cell Migration.

Prolactin (PRL) is a polypeptide hormone synthesized in the lactotrophs of the adenohypophysis and in extrahypophyseal glands (such as the prostate and breasts) where it promotes their development. PRL is also involved in cancer development in these glands. It has been shown to stimulate cancer cell migration, suggesting its possible involvement in metastasis, in which cell migration plays an essential role. However, the role of PRL in cell migration is still unclear. Moreover, the intracellular mechanisms activated by PRL to carry out cell migration are less well understood. PRL exerts its effects via the PRL receptor (PRLR), which leads intracellularly to phosphorylation of Janus protein kinase 2 (JAK2), which in turn phosphorylates p21-activated protein kinase (PAK1), leading to an increase in cell migration. Although several studies have described the involvement of the PRL-PAK1 pathway in breast cancer cell migration, the molecular mechanisms have not been fully elucidated and there is no integration of these into signaling pathways. This study was conducted based on literature search of review articles and original research in the PubMed database, using the following keywords: PRL, cell migration, PRL and cell migration, PAK1 and signaling pathways. The aim of this review article was to describe the major signaling pathways controlled by PRL-PAK1 and propose a comprehensive model of the signaling pathways associated with PRL-PAK1.

Antioxidant Activity and Kinetic Characterization of Chlorella vulgaris Growth under Flask-Level Photoheterotrophic Growth Conditions.

C. vulgaris is a unicellular microalgae, whose growth depends on the conditions in which it is found, synthesizing primary and secondary metabolites in different proportions. Therefore, we analyzed and established conditions in which it was possible to increase the yields of metabolites obtained at the flask level, which could then be scaled to the photobioreactor level. As a methodology, a screening design was applied, which evaluated three factors: type of substrate (sodium acetate or glycerol); substrate concentration; and exposure-time to red light (photoperiod: 16:8 and 8:16 light/darkness). The response variables were: cell division; biomass; substrate consumption; and antioxidant activity in intracellular metabolites (ABTS•+ and DPPH•). As a result, the sodium acetate condition of 0.001 g/L, in a photoperiod of 16 h of light, presented a doubling time (Td = 4.84 h) and a higher rate of division (σ = 0.20 h-1), having a final biomass concentration of 2.075 g/L. In addition, a higher concentration of metabolites with antioxidant activity was found in the sodium acetate (0.629 Trolox equivalents mg/L ABTS•+ and 0.630 Trolox equivalents mg/L DPPH•). For the glycerol, after the same photoperiod (16 h of light and 8 h of darkness), the doubling time (Td) was 4.63 h, with a maximum division rate of σ = 0.18 h-1 and with a biomass concentration at the end of the kinetics of 1.4 g/L. Sodium acetate under long photoperiods, therefore, is ideal for the growth of C. vulgaris, which can then be scaled to the photobioreactor level.

The mobility of thorium, uranium and rare earth elements from Mid Ordovician black shales to acid waters and its removal by goethite and schwertmannite.

Previous studies have addressed the occurrence of Acid Rock Drainage (ARD) affecting La Silva stream due to the generation of large dumps of Middle Ordovician black shales during the construction of a highway close to El Bierzo (León, Spain). This ARD was characterized by sulphated acid waters with high concentration of heavy metals and anomalies in dissolved thorium (Th) and uranium (U). In the present study, we analyse in depth black shales and water, streambed sediments and precipitates of La Silva stream and its tributaries using different petrographic, mineralogical and geochemical approaches. Black shales, with average Th and U contents of 20 and 3 µg/g respectively contain disseminated detritic micro-grains of high weathering-resistant minerals, such as monazite and xenotime, that present smaller amounts of yttrium and rare earth elements (REY) and other elements as Ca, U, Th, Si and F. Results of the affected waters by ARD show an enrichment in dissolved Th, U and REY of several orders of magnitude with respect to natural waters. Sampled precipitates were mainly schwertmannite (Fe8O8(OH)8-2x (SO4)xO16•nH2O) and goethite (α-Fe3+O(OH)) that showed an enrichment of Th (up to 798 µg/g) and REY, due to the presence of dissolved anionic species (e.g. [Formula: see text] , [Formula: see text] ) that enables their adsorption. Furthermore, these black shales show a clear enrichment in REE (Rare Earth Elements) with respect to NASC (North American Shales Composite) normalized REE patterns. Likewise, normalized REE patterns of stream waters and precipitates clearly show convex curvatures in middle-REE (MREE) with respect to light- and something less than heavy-REE, indicating the trend towards MREE enrichment. These findings are essential to evaluate the impact of ARD of Mid Ordovician shales in the surrounding environment, and to start considering these site as potential source of REE and critical raw materials, activating a Circular Economy.

Characterization of Candida albicans and Staphylococcus aureus polymicrobial biofilm on different surfaces.

BACKGROUND: Staphylococcus aureus and Candida albicans have been co-isolated from biofilm-associated diseases such as denture stomatitis, periodontitis, and burn wound infections, as well as from medical devices. However, the polymicrobial biofilm of both microorganisms has not been fully characterized. AIMS: To characterize the polymicrobial biofilm of C. albicans and S. aureus in terms of microbial density, synergy, composition, structure, and stability against antimicrobials and chemical agents. METHODS: Crystal violet assay was used to measure the biofilm formation. Scanning electron microscopy and confocal microscopy were used to analyze the structure and chemical composition of the biofilms, respectively. RESULTS: Supplemented media with fetal bovine serum (FBS) decreased the biofilm formation of S. aureus and the polymicrobial biofilm. For C. albicans, depending on the culture media, the addition of glucose or FBS had a positive effect in biofilm formation. FBS decreased the adhesion to polystyrene wells for both microorganisms. Supplementing the media with glucose and FBS enhanced the growth of C. albicans and S. aureus, respectively. It seems that C. albicans contributes the most to the adhesion process and to the general structure of the biofilms on all the surfaces tested, including a catheter model. Interestingly, S. aureus showed a great adhesion capacity to the surface of C. albicans in the biofilms. Proteins and ß-1,6-linked polysaccharides seem to be the most important molecules in the polymicrobial biofilm. CONCLUSIONS: The polymicrobial biofilm had a complex structure, with C. albicans serving as a scaffold where S. aureus adheres, preferentially to the hyphal form of the fungus. Detection of polymicrobial infections and characterization of biofilms will be necessary in the future to provide a better treatment.

Characterization of Candida albicans and Staphylococcus aureus polymicrobial biofilm on different surfaces

Background:Staphylococcus aureus and Candida albicans have been co-isolated from biofilm-associated diseases such as denture stomatitis, periodontitis, and burn wound infections, as well as from medical devices. However, the polymicrobial biofilm of both microorganisms has not been fully characterized.Aims:To characterize the polymicrobial biofilm of C. albicans and S. aureus in terms of microbial density, synergy, composition, structure, and stability against antimicrobials and chemical agents.Methods:Crystal violet assay was used to measure the biofilm formation. Scanning electron microscopy and confocal microscopy were used to analyze the structure and chemical composition of the biofilms, respectively.Results:Supplemented media with fetal bovine serum (FBS) decreased the biofilm formation of S. aureus and the polymicrobial biofilm. For C. albicans, depending on the culture media, the addition of glucose or FBS had a positive effect in biofilm formation. FBS decreased the adhesion to polystyrene wells for both microorganisms. Supplementing the media with glucose and FBS enhanced the growth of C. albicans and S. aureus, respectively. It seems that C. albicans contributes the most to the adhesion process and to the general structure of the biofilms on all the surfaces tested, including a catheter model. Interestingly, S. aureus showed a great adhesion capacity to the surface of C. albicans in the biofilms. Proteins and β-1,6-linked polysaccharides seem to be the most important molecules in the polymicrobial biofilm. (AU)

Antecedentes:Staphylococcus aureus y Candida albicans son aislados conjuntamente de infecciones asociadas a la formación de biopelículas, tales como periodontitis, estomatitis e infecciones provenientes de quemaduras, así como en dispositivos médicos. Sin embargo, la biopelícula formada por ambos microorganismos no ha sido completamente caracterizada.Objetivos:Caracterizar la biopelícula de C. albicans y S. aureus en cuanto a densidad microbiana, sinergismo, composición, estructura y estabilidad frente a agentes químicos y antimicrobianos.Métodos:El análisis de la formación de biopelícula se realizó mediante el ensayo de cristal violeta. Se analizó la composición química y la estructura de las biopelículas mediante microscopio confocal y microscopio electrónico de barrido, respectivamente.Resultados:La adición al medio de suero bovino fetal (SBF) redujo la biopelícula mono- y polimicrobiana de S. aureus. En C. albicans, con la adición de glucosa o SBF, se incrementó la formación de biopelícula. La adhesión de los microorganismos a las placas de poliestireno se redujo en presencia de SBF. La suplementación del medio con glucosa y SBF favoreció la proliferación de C. albicans y S. aureus, respectivamente. C. albicans mostró una mejor adhesión y contribuyó más a la densidad total de la biopelícula en diferentes superficies probadas, incluyendo un modelo de catéter. De manera interesante, S. aureus mostró una mejor adhesión a la superficie de C. albicans en la biopelícula. Las proteínas y los polisacáridos con enlaces β-1,6 parecen ser las moléculas más abundantes en la biopelícula. (AU)

An in vitro study of ApxI from Actinobacillus pleuropneumoniae serotype 10 and induction of NLRP3 inflammasome-dependent cell death.

BACKGROUND: Actinobacillus pleuropneumoniae (AP) is the causative agent of porcine pleuropneumonia. Apx exotoxins are the most important virulence factors associated with the induction of lesions. ApxI is highly cytotoxic on a wide range of cells. Besides the induction of necrosis and apoptosis of ApxI on porcine alveolar macrophages (PAMs), its role in pyroptosis, a caspase-1-dependent form of cell death, has not been reported. The aim of this study was to analyse if NLRP3 inflammasome participates in cell death induced by ApxI. METHODS: PAMs, the porcine alveolar macrophage cell line 3D4/21 and a porcine aortic endothelial cell line were used in this study. We used Z-VAD-FMK and Ac-YVAD-cmk to inhibit caspase-1. Glyburide and MCC950 were used to inhibit the NLRP3 inflammasome. A lactate dehydrogenase release assay was used to measure the percentage of cell death. Caspase-1 expression was analysed by immunofluorescence. End-point RT-PCR was used to analyse the expression of NLRP3 mRNA. RESULTS: Rapid cell death in PAMs, 3D4/21 cells and the endothelial cell line were induced by ApxI. This cell death decreased by using caspase-1 and NLRP3 inflammasome inhibitors and by blocking the K+ efflux. Expression of NLRP3 mRNA was induced by ApxI in alveolar macrophages while it was constitutive in the endothelial cell line. Detection of caspase-1 in alveolar macrophages was higher after ApxI treatment and it was blocked by MCC950 or heat inactivation. CONCLUSIONS: To the best of the authors' knowledge, we have described for the first time in vitro induction of ApxI associated pyroptosis in alveolar macrophages and endothelial cells, a rapid cell death that depends on the activation of caspase-1 via the NLRP3 inflammasome.

Insights into Growth Factors in Liver Carcinogenesis and Regeneration: An Ongoing Debate on Minimizing Cancer Recurrence after Liver Resection.

Hepatocellular carcinoma has become a leading cause of cancer-associated mortality throughout the world, and is of great concern. Currently used chemotherapeutic drugs in the treatment of hepatocellular carcinoma lead to severe side effects, thus underscoring the need for further research to develop novel and safer therapies. Liver resection in cancer patients is routinely performed. After partial resection, liver regeneration is a perfectly calibrated response apparently sensed by the body's required liver function. This process hinges on the effect of several growth factors, among other molecules. However, dysregulation of growth factor signals also leads to growth signaling autonomy and tumor progression, so control of growth factor expression may prevent tumor progression. This review describes the role of some of the main growth factors whose dysregulation promotes liver tumor progression, and are also key in regenerating the remaining liver following resection. We herein summarize and discuss studies focused on partial hepatectomy and liver carcinogenesis, referring to hepatocyte growth factor, insulin-like growth factor, and epidermal growth factor, as well as their suitability as targets in the treatment of hepatocellular carcinoma. Finally, and given that drugs remain one of the mainstay treatment options in liver carcinogenesis, we have reviewed the current pharmacological approaches approved for clinical use or research targeting these factors.

The Periaqueductal Gray and Its Extended Participation in Drug Addiction Phenomena.

The periaqueductal gray (PAG) is a complex mesencephalic structure involved in the integration and execution of active and passive self-protective behaviors against imminent threats, such as immobility or flight from a predator. PAG activity is also associated with the integration of responses against physical discomfort (e.g., anxiety, fear, pain, and disgust) which occurs prior an imminent attack, but also during withdrawal from drugs such as morphine and cocaine. The PAG sends and receives projections to and from other well-documented nuclei linked to the phenomenon of drug addiction including: (i) the ventral tegmental area; (ii) extended amygdala; (iii) medial prefrontal cortex; (iv) pontine nucleus; (v) bed nucleus of the stria terminalis; and (vi) hypothalamus. Preclinical models have suggested that the PAG contributes to the modulation of anxiety, fear, and nociception (all of which may produce physical discomfort) linked with chronic exposure to drugs of abuse. Withdrawal produced by the major pharmacological classes of drugs of abuse is mediated through actions that include participation of the PAG. In support of this, there is evidence of functional, pharmacological, molecular. And/or genetic alterations in the PAG during the impulsive/compulsive intake or withdrawal from a drug. Due to its small size, it is difficult to assess the anatomical participation of the PAG when using classical neuroimaging techniques, so its physiopathology in drug addiction has been underestimated and poorly documented. In this theoretical review, we discuss the involvement of the PAG in drug addiction mainly via its role as an integrator of responses to the physical discomfort associated with drug withdrawal.

Influence of hail suppression systems over silver content in the environment in Aragón (Spain). I: Rainfall and soils.

In several countries, hail is considered as the most harmful climatic phenomenon from an agricultural perspective. The surroundings of the Gallocanta Lake (North-East Spain), is one of the areas where the storms affecting the Ebro Valley are formed. For this reason, silver iodide from hail suppression systems has been emitted to the atmosphere for half a century. Nowadays, there is an increasing social concern about the potential environmental consequences of this activity, which has promoted the analysis of the influence of hail suppression systems regarding the amount of silver concentration in the ecosystem. This study focuses on silver atmospheric deposition and its accumulation in soils. To this end, silver concentrations in rainfall (5 gauges, 16 samples per site, from April 2017 to March 2019) and soils (72 samples) distributed across the hail suppression network managed by the Anti-hail Consortium of Aragón, were analysed. The results show that the amount of silver is much higher in rainfall gauges and soils close to ground-based silver iodide generators (85 µg/m2·day and 10 mg/kg soil, respectively), but concentrations considerably decrease when samples are collected far from them (downing to 0.3 µg/m2·day and 0.1 mg/kg soil). Apart from the samples obtained nearby silver iodide generators, most of the other soil samples display silver concentrations below the legal threshold established for the most vulnerable activities (1 mg/kg soil in agricultural and forestry land uses). Nevertheless, silver content, both in precipitation and in soils, is higher in regions where hail suppression has been developed for decades when compared to nearby areas in which silver iodide emissions did not occur. Silver content observed in soils is not high, but their cumulative effect in sediments and biota should be analysed, which is the aim of the second part of the present study.

1-Boc-Piperidine-4-Carboxaldehyde Prevents Binge-Eating Behaviour and Anxiety in Rats.

BACKGROUND: Piperidines are biogenic amines studied mainly in toxicology because they were initially found as alkaloids from peppers and insect venoms. Piperidines are also produced in the human body, and their actions seem to be related to wakefulness/sleep and other cognitive phenomena. Piperidines have been minimally characterized for therapeutic applications. In this context, 1-Boc-piperidine-4-carboxaldehyde (1-Boc-piperidine) is a piperidine-derivative molecule with no mechanism of action reported, although its uses include the synthesis of GPR119 selective agonists that have been patented as anti-obesity drugs. OBJECTIVES: The aim of this work was to study the effects of 1-Boc-piperidine on binge-eating behaviour and anxiety in Wistar rats. METHODS: In experimental protocol 1, binge-eating behaviour was induced in animals that received pre-treatment (i.p.) with (i) vehicle (methanol 10%; 1 mL/kg), (ii) 1-Boc-piperidine (1 µmol kg-1), or (iii) 1-Boc-piperidine (10 µmol kg-1). In experimental protocol 2, mildly stressed animals were evaluated in the elevated plus maze under the acute effects of the pre-treatments applied in experimental protocol 1. RESULTS AND CONCLUSIONS: 1-Boc-piperidine decreased, in a dose-dependent manner, the intake of calories from a succulent hyper-caloric food in a binge-eating protocol in female rats, whereas the acute exposition to this piperidine exerted an anxiolytic effect in the male rat. In both effects, the mechanism of action remains to be characterized.

Susceptibility for Some Infectious Diseases in Patients With Diabetes: The Key Role of Glycemia.

Uncontrolled diabetes results in several metabolic alterations including hyperglycemia. Indeed, several preclinical and clinical studies have suggested that this condition may induce susceptibility and the development of more aggressive infectious diseases, especially those caused by some bacteria (including Chlamydophila pneumoniae, Haemophilus influenzae, and Streptococcus pneumoniae, among others) and viruses [such as coronavirus 2 (CoV2), Influenza A virus, Hepatitis B, etc.]. Although the precise mechanisms that link glycemia to the exacerbated infections remain elusive, hyperglycemia is known to induce a wide array of changes in the immune system activity, including alterations in: (i) the microenvironment of immune cells (e.g., pH, blood viscosity and other biochemical parameters); (ii) the supply of energy to infectious bacteria; (iii) the inflammatory response; and (iv) oxidative stress as a result of bacterial proliferative metabolism. Consistent with this evidence, some bacterial infections are typical (and/or have a worse prognosis) in patients with hypercaloric diets and a stressful lifestyle (conditions that promote hyperglycemic episodes). On this basis, the present review is particularly focused on: (i) the role of diabetes in the development of some bacterial and viral infections by analyzing preclinical and clinical findings; (ii) discussing the possible mechanisms by which hyperglycemia may increase the susceptibility for developing infections; and (iii) further understanding the impact of hyperglycemia on the immune system.

Razor scrape assay, an alternative variation to wound and healing assays.

Cell migration is the process by which cells move through tissues, and it is crucial to carry out a wide variety of physiological and pathological processes. The study methods to evaluate cell migration are very useful tools for biomedical research. Among these methods, the wound and healing assay is one of the simplest, most economical and is widely used in research. However, one of its disadvantages is that the width and shape of the wound can vary among experimental samples since the scraping is carried out manually, representing a difficult variable to control. In the present article a variant of the razor scrape assay is addressed, which eliminates this variation in the width of the wound, thus facilitating the measurement and comparison using the total area of cell migration.•A method that can be carried out under standard culture conditions.•Avoids the disadvantage of variation in width and shape of the wound.•It constitutes a simple, cheap option and multiple advantages over the traditional method.

NPY-Y1 receptors in dorsal periaqueductal gray modulate anxiety, alcohol intake, and relapse in Wistar rats.

Neuropeptide Y (NPY) is likely the main endogenous anxiolytic neuromodulator involved in alcohol intake. NPY-Y1, a receptor for NPY, is highly expressed in the periaqueductal gray (PAG), a mesencephalic structure involved in integrating nervous activity to the performance of active and passive defensive behaviors related to fear and anxiety. Interestingly, anxiety and fear are some of the prevailing emotional negative states during alcohol abstinence. Moreover, an inverse relationship between NPY activity and alcohol consumption has been frequently reported, mainly in the extended amygdala. Nevertheless, both the roles of NPY and that of the receptor involved in these actions have been scarcely studied. Thus, the aim of this study was to analyze the pharmacological effect of NPY and NPY-Y1 receptor blockade into the dorsal periaqueductal gray (D-PAG) in an alcohol consumption and relapse paradigm in adult male Wistar rats. Ninety-six rats at postnatal day 42 (PND-42) were classified as having low and high anxiety (LA and HA), respectively, through the elevated plus maze test (EPM). Then, those animals were randomly divided into alcohol naïve (AN) and forced alcohol consumption (FAC) groups. A cannula was implanted in D-PAG to microinject vehicle (VEH), NPY, or BIBP-3226 (a selective NPY-Y1 receptor antagonist). A defensive burying behavior test (DBB) was performed to assess the anxiety-like state during withdrawal, followed by a 24-hour free choice voluntary alcohol intake test. Under our experimental conditions, NPY microinjection decreased alcohol consumption in HA rats, whereas NPY-Y1 receptor blockade in D-PAG produced a notably anxiogenic effect and higher alcohol intake and relapse. In conclusion, NPY in the D-PAG, most likely acting on NPY-Y1 receptors, induced a significant anxiolytic effect and prominently inhibited alcohol consumption and relapse in Wistar rats.

Phenomenology and geographical gradients of atmospheric deposition in southwestern Europe: Results from a multi-site monitoring network.

This article presents the results of atmospheric deposition from a 15-sites network which cover remote, agricultural, urban and industrial areas in the Iberian Peninsula and the Balearic Islands, with the aim of exploring geographical, climatic and natural vs anthropogenic gradients. Annual average fluxes of global deposition, discriminating insoluble (3,5-20,7 g m-2 year-1) and soluble-inorganic (7,1-45,5 g m-2 year-1) aerosols are discussed, seasonal patterns are regarded, and an attempt to estimate the impact of the main sources is presented. The wide range of atmospheric deposition fluxes (DF) regarding soluble (DFSOL) and insoluble (DFINS) has been investigated taking into consideration the contribution from nearby to long-distance sources, such as African dust, or regional-to-nearby ones, which include agricultural dust in the Ebro Valley, industrial emissions at different parts, urban dust at all cities, or saline dust resuspension from a dissicated lake bed. DFSOL is made up of marine aerosols, prevailing in coastal areas, with few exceptions in the Ebro Valley; nitrogen-species, homogeneously distributed across the network, with few exceptions due to agricultural sources; mineral dust, enhanced in the Ebro Valley owing to regional and agricultural emissions; and phospathe, displaying comparable values to other studies in general, but three hotspots at regional background environments have been identified. DFINS particles followed the aridity pattern, especially where anthropogenic emissions take place. Our estimates indicate that the regional dust to DFINS in the Ebro Valley represented 23-30%, overpassing 50% at intensive agricultural areas. Similarly, urban-metropolitan contributions accounted for 37-45% at the four cities, and 55% at the industrial one. African dust deposition was enhanced in the Central Pyrenees (75-80%) as a result of the magnification of atmospheric washout processes, and in south-eastern Iberia (69%) owing to the higher frequency of dust outbreaks.