Inflammatory and non-inflammatory monocytes as novel prognostic biomarkers of survival in SOD1G93A mouse model of Amyotrophic Lateral Sclerosis

Amyotrophic Lateral Sclerosis (ALS) has lately become a suitable scenario to study the interplay between the hematopoietic system and disease progression. Recent studies in C9orf72 null mice have demonstrated that C9orf72 is necessary for the normal function of myeloid cells. In this study, we aimed to analyze in depth the connection between the hematopoietic system and secondary lymphoid (spleen) and non-lymphoid (liver and skeletal muscle) organs and tissues along the disease progression in the transgenic SOD1G93A mice. Our findings suggested that the inflammatory response due to the neurodegeneration in this animal model affected all three organs and tissues, especially the liver and the skeletal muscle. However, the liver was able to compensate this inflammatory response by means of the action of non-inflammatory monocytes, while in the skeletal muscle inflammatory monocytes prompted a further inflammation process until the terminal state of the animals. Interestingly, in blood, a positive correlation was found between non-inflammatory monocytes and survival of the transgenic SOD1G93A mice, while the contrary (a negative correlation) was found in the case of inflammatory monocytes, supporting their potential role as biomarkers of disease progression and survival in this animal model. These findings could prompt future translational studies in ALS patients, promoting the identification of new reliable biomarkers of disease progression

Effects of the 2021 La Palma volcanic eruption on groundwater resources (part II): Hydrochemical impacts

Volcanic eruptions can have significant implications for the management and sustainability of water resources in volcanic islands. The recent 2021 eruption of the Tajogaite volcano in La Palma Island (Canary Islands, Spain) raised concerns regarding its potential impact on groundwater resources. This study is the second part of a series investigating the hydrogeochemical impacts of the eruption. The study involved conducting three groundwater sampling campaigns during the eruption and two after the eruption, six months and one year after the eruption ceased. A total of 15 monitored points, including piezometers, wells, water galleries, and main gully collector of the island, all relatively close (2–15 km) to the erupted volcano, were sampled for the analysis of major, minor, and trace elements, physiochemical parameters, which were measured on-site. Statistical analyses were performed to assess the differences in groundwater composition before, during, and after the eruption. To evaluate the differences in water quality compared to pre-eruption events, 33 additional historical groundwater samples provided by the local Water Authority were assessed, and 103 groundwater analysis results from the groundwater data base of the Spanish National Geological Survey (IGME) were considered. The results of the study showed low but statistically significant changes in pH, T, conductivity and groundwater composition, mainly related to the high increases in several trace element concentrations, such as Al, Cr, Fe, Mo, Ni, Sr, Th, Tl, V, Zn, Ba, Cd, Co, Cu, Pb and U, with increments in various orders of magnitude for several elements. This increase was found to be highly influenced by the sample distance to the volcano during the eruption stage. The significance of these findings lies in their usefulness to enhance our understanding of the effects of volcanic eruptions on groundwater quality resources and demonstrate their resilience to this hazardous phenomenon, which ultimately underscores their reliability

A Review on the Current Knowledge on ZIKV Infection and the Interest of Organoids and Nanotechnology on Development of Effective Therapies against Zika Infection

Zika virus (ZIKV) acquired a special relevance due to the pandemic that occurred in the Americas in 2015, when an important number of fetal microcephaly cases occurred. Since then, numerous studies have tried to elucidate the pathogenic mechanisms and the potential therapeutic approaches to combat the virus. Cellular and animal models have proved to be a basic resource for this research, with the more recent addition of organoids as a more realistic and physiological 3D culture for the study of ZIKV. Nanotechnology can also offer a promising therapeutic tool, as the nanoparticles developed by this field can penetrate cells and deliver a wide array of drugs in a very specific and controlled way inside the cells. These two state-of-the-art scientific tools clearly provide a very relevant resource for the study of ZIKV, and will help researchers find an effective treatment or vaccine against the virus

Autofagia y apoptosis: Mecanismos de regulación celular en un modelo murino de ELA

La Esclerosis Lateral Amiotrófica (ELA) es una enfermedad neurodegenerativa progresiva y fatal, caracterizada por una degeneración de las neuronas motoras y una atrofia muscular y posterior parálisis muscular, que desembocan la muerte del paciente, normalmente por parada respiratoria, entre los 2 y 5 años desde el inicio de los síntomas. El objetivo fundamental del presente trabajo se basó en el estudio de los niveles de expresión proteicos de marcadores de apoptosis (BAX, BCL-2, CSP3 y PROCSP3) y de autofagia (ATG5, BECLIN1, E2F1 y LC3) en músculo esquelético del modelo murino SOD1G93A en los tres estadios principales de la enfermedad. Los resultados mostraron una activación significativa de los niveles de expresión de todos los marcadores de apoptosis estudiados en el estadio sintomático temprano. Sin embargo, en el estadio sintomático, no se observaron niveles significativamente elevados de ningún marcador de autofagia ni de apoptosis. Finalmente, en el estadio terminal, todos los marcadores de autofagia mostraron un incremento significativo en sus niveles de expresión. Estos resultados parecen indicar que la apoptosis se encuentra activada en etapas tempranas de la enfermedad, mientras que la autofagia comienza a activarse en etapas más tardías. Los niveles de expresión que se registraron para la proteína p62, que está estrechamente relacionada con el sistema de degradación proteíco ubiquitina-proteasoma (UPS), autofagia (macroautofagia) e incluso con la apoptosis, parecen indicar que en el estadio central de la enfermedad, en el que no se observa una activación de ningún marcador de autofagia ni de apoptosis, podría jugar un papel relevante la activación del UPS. Con estos resultados, podemos concluir que tanto la apoptosis como la autofagia participan en la regulación de la muerte celular en el músculo esquelético de animales transgénicos SOD1G93A modelo para la ELA, siendo necesarios estudios posteriores para determinar si la modulación de estos procesos podría servir como diana terapéutica

Input Rate (IR) differences between WT and SOD1G93A mice along time.

<p>Each IR represents the difference between the cell percentage at one given time point and the preceding one, divided by the days elapsed between the two blood extractions. <i>y</i>-axis represents the mean cell percentage released per day for each cell type studied, while <i>x</i>-axis represents each IR. Control mice are represented with rhombuses and SOD1G93A mice with squares. The IR was used to assess the release efficiency of <b>(A)</b> HSCs, <b>(B)</b> CLPs and <b>(C)</b> CMPs to the blood stream. IR values are represented as percentage means per day ± s.e.m. Asterisks represent an independent t-test <i>p</i>-value < 0.05 (*); n = 12 animals per genotype (extractions were performed serially along time, always using the same mice).</p

RT-PCR and Western blot study of monocytic markers CCR2, CX3CR1 and Ly6C in the liver of WT and transgenic SOD1G93A mice.

<p><b>(A, B, C)</b> RT-PCR was used to measure mRNA expression levels of <i>Ccr2</i>, expressed in inflammatory monocytes; <i>Cx3cr1</i>, a marker of non-inflammatory monocytes; and <i>Ly6C</i>, highly expressed in inflammatory monocytes. <b>(D, E, F)</b> Western blot analysis of CCR2, CX3CR1 and Ly6C protein expression levels. <i>y</i>-axis and <i>x</i>-axis represent the gene expression fold change and time points selected for RT-PCR analysis, and protein fold change and time point selected for Western Blot analysis, respectively. Transgenic SOD1G93A mice are represented with black bars and WT litter mates used as controls, with light grey bars. Error bars represent the standard error of the mean (s.e.m.) for the Western blot analysis and error bars for the RT-PCR analysis were calculated using the method developed by Livak et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0184626#pone.0184626.ref030" target="_blank">30</a>]. Statistical significance was established at <i>p</i>-values under 0.05 (*) and high statistical significance at <i>p</i>-values under 0.01 (**). n = 12 animals per genotype and time point. <b>(G)</b> Representative protein bands for every protein studied by Western blot in both WT and SOD1G93A mice. GAPDH was used as housekeeping protein for protein normalization.</p

RT-PCR and Western blot study of monocytic markers CCR2, CX3CR1 and Ly6C in the skeletal muscle of WT and transgenic SOD1G93A mice.

<p><b>(A, B, C)</b> RT-PCR was used to measure mRNA expression levels of <i>Ccr2</i>, expressed in inflammatory monocytes; <i>Cx3cr1</i>, a marker of non-inflammatory monocytes; and <i>Ly6C</i>, expressed in inflammatory monocytes. <b>(D, E, F)</b> Western blot analysis of CCR2, CX3CR1 and Ly6C protein expression levels. <i>y</i>-axis and <i>x</i>-axis represent the gene expression fold change and time points selected for RT-PCR analysis, and protein fold change and time point selected for Western Blot analysis, respectively. Transgenic SOD1G93A mice are represented with black bars and WT litter mates used as controls, with light grey bars. Error bars represent the standard error of the mean (s.e.m.) for the Western blot analysis and error bars for the RT-PCR analysis were calculated using the method developed by Livak et al. [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0184626#pone.0184626.ref030" target="_blank">30</a>]. Statistical significance was established at <i>p</i>-values under 0.05 (*) and high statistical significance at <i>p</i>-values under 0.01 (**). n = 12 animals per genotype and time point. <b>(G)</b> Representative protein bands for every protein studied by Western blot in both WT and SOD1G93A mice. GAPDH was used as housekeeping protein for protein normalization.</p

Potential role of monocytes in the survival of transgenic SOD1G93A mice.

<p><b>(A)</b> Total monocytes, <b>(B)</b> inflammatory and <b>(C)</b> non-inflammatory monocytes percentages were measured by flow cytometry in the blood of WT (light grey bars) and transgenic SOD1G93A mice (black bars) at the main stages of the disease: asymptomatic stage (P50), symptomatic stage (P75) and terminal stage (mice endpoint ranged between 128 and 143 days, depending on the survival of the individual animals). WT litter mates were used as controls. <i>y</i>-axis represents the mean cell percentage for each cell type studied, while <i>x</i>-axis represents the time points selected for blood extraction. The gating strategy of the flow cytometry analysis is shown in <b>(D)</b>. Finally, the correlations found between monocyte types and survival is shown in <b>(E)</b>. Data are presented as means and error bars represent the standard error of the mean (s.e.m.). Statistical significance was established at <i>p</i>-values under 0.05 (*). n = 12 animals per genotype (extractions were performed serially along time, always using the same mice).</p

Decreased waterborne pathogenic bacteria in an urban aquifer related to intense shallow geothermal exploitation

The implications of intensive use of shallow geothermal energy resources in shallow urban aquifers are still not known for waterborne pathogens relevant to human health. Firstly, we hypothesized that waterborne enteric pathogens would be relatively increased in heated groundwater plumes. To prove this, microbiological sampling of 31 piezometers covering the domain of an urban groundwater body affected by microbiological contamination and energetically exploited by 70 groundwater heat pump systems was performed. Mean differences of pathogenic bacteria contents between impacted and non-impacted monitoring points were assessed with a two-tailed independent Student's t-test or Mann-Whitney U and correlation coefficients were also calculated. Surprisingly, the results obtained revealed a significant and generalized decrease in waterborne pathogen contents in thermally impacted piezometers compared to that of nonimpacted piezometers. This decrease is hypothesized to be caused by a heat shock to bacteria within the heat exchangers. The statistically significant negative correlations obtained between waterborne pathogen counts and temperature could be explained by the spatial distribution of the bacteria, finding that bacteria start to recover with increasing distance from the injection point. Also, different behavior groups fitting exponential regression models were found for the bacteria species studied, justified by the different presence and influence of several aquifer parameters and major, minor and trace elements studied, as well as the coexistence with other bacteria species. The results obtained from this work reinforce the concept of shallow geothermal resources as a clean energy source, as they could also provide the basis to control the pathogenic bacteria contents in groundwater bodies. (C) 2018 Elsevier B.V. All rights reserved

HSPCs percentages recorded by flow cytometry in the spleen of WT and transgenic SOD1G93A mice.

<p><b>(A)</b> HSC, <b>(B)</b> CLP and <b>(C)</b> CMP percentages were measured by flow cytometry in the spleen of transgenic SOD1G93A mice (black bars) at the main stages of the disease: asymptomatic stage (P50), symptomatic stage (P75) and terminal stage (sacrifice). WT litter mates (light grey bars) were used as controls. The gating strategy of the flow cytometry analysis is shown in <b>(D)</b>. <i>y</i>-axis represents the mean cell percentage for each cell type, and <i>x</i>-axis represents the time points selected. Data are represented as means and error bars represent the standard error of the mean (s.e.m.). Statistical significance was established at <i>p</i>-values under 0.05 (*), high statistical significance at <i>p</i>-values under 0.01 (**) and very high statistical significance at <i>p</i>-values under 0.001 (***). n = 12 animals per genotype and time point.</p