Mining threats in high-level biodiversity conservation policies.

Amid a global infrastructure boom, there is increasing recognition of the ecological impacts of the extraction and consumption of construction minerals, mainly processed as concrete, including significant and expanding threats to global biodiversity. We investigated how high-level national and international biodiversity conservation policies address mining threats, with a special focus on construction minerals. We conducted a review and quantified the degree to which threats from mining these minerals are addressed in biodiversity goals and targets under the 2011-2020 and post-2020 biodiversity strategies, national biodiversity strategies and action plans, and the assessments of the Intergovernmental Science-Policy Platform on Biodiversity and Ecosystem Services. Mining appeared rarely in national targets but more frequently in national strategies. Yet, in most countries, it was superficially addressed. Coverage of aggregates mining was greater than coverage of limestone mining. We outline 8 key components, tailored for a wide range of actors, to effectively mainstream biodiversity conservation into the extractive, infrastructure, and construction sectors. Actions include improving reporting and monitoring systems, enhancing the evidence base around mining impacts on biodiversity, and modifying the behavior of financial agents and businesses. Implementing these measures could pave the way for a more sustainable approach to construction mineral use and safeguard biodiversity.

Amenazas de la minería a las políticas de alto nivel para la conservación de la biodiversidad Resumen Enmedio del auge global del desarrollo de infraestructura, hay un mayor reconocimiento de los impactos ecológicos de la extracción y consumo de materiales para construcción, procesados predominantemente como concreto. Estos materiales representan amenazas significativas y en expansión para la biodiversidad global. Investigamos cómo son abordadas las amenazas de la minería por las políticas nacionales e internacionales de alto nivel para la conservación de la biodiversidad, con enfoque especial en los minerales para construcción. Realizamos una revisión exhaustiva y cuantificamos el grado en el cual son abordadas las amenazas de la extracción de estos minerales en los objetivos y metas para la biodiversidad bajo estrategias 2011­2020 y post 2020, las estrategias y planes de acción nacionales para la biodiversidad, y las evaluaciones de la Plataforma Intergubernamental Científico­normativa sobre Diversidad Biológica y Servicios de los Ecosistemas. La minería raramente apareció en los objetivos nacionales, pero fue más frecuente en las estrategias nacionales. Sin embargo, fue abordada superficialmente en la mayoría de los países. La cobertura de minería de agregados fue mayor que la cobertura de la minería de caliza. Describimos ocho componentes clave, adaptados para una amplia gama de actores, para incorporar eficazmente la conservación de la biodiversidad en los sectores extractivo, desarrollo de infraestructura y construcción. Las acciones incluyen la mejora de los sistemas de informes y monitoreo, el reforzamiento de la base de evidencias en torno a los impactos de la minería sobre la biodiversidad y la modificación del comportamiento de los agentes financieros y comerciales. La implementación de estas medidas podría allanar el camino para un enfoque más sostenible en el uso de minerales para la construcción y la salvaguarda de la biodiversidad.

Anthelmintic efficacy of an organic fraction from Guazuma ulmifolia leaves and evaluation of reactive oxidative stress on Haemonchus contortus.

This study describes the anthelmintic efficacy of an organic fraction (EtOAc-F) from Guazuma ulmifolia leaves and the evaluation of its reactive oxidative stress on Haemonchus contortus. The first step was to assess the anthelmintic effect of EtOAc-F at 0.0, 3.5, 7.0 and 14 mg kg of body weight (BW) in gerbil's (Meriones unguiculatus) artificially infected with H. contortus infective larvae (L3). The second step was to evaluate the preliminary toxicity after oral administration of the EtOAc-F in gerbils. Finally, the third step was to determine the relative expression of biomarkers such as glutathione (GPx), catalase (CAT), and superoxide dismutase (SOD) against H. contortus L3 post-exposition to EtOAc-F. Additionally, the less-polar compounds of EtOAc-F were identified by gas mass spectrophotometry (GC-MS). The highest anthelmintic efficacy (97.34%) of the organic fraction was found in the gerbils treated with the 14 mg/kg of BW. Histopathological analysis did not reveal changes in tissues. The relative expression reflects overexpression of GPx (p<0.05, fold change: 14.35) and over expression of SOD (p≤0.05, fold change: 0.18) in H. contortus L3 exposed to 97.44 mg/mL of EtOAc-F compared with negative control. The GC-MS analysis revealed the presence of 4-hydroxybenzaldehyde (1), leucoanthocyanidin derivative (2), coniferyl alcohol (3), ferulic acid methyl ester acetate (4), 2,3,4-trimethoxycinnamic acid (5) and epiyangambin (6) as major compounds. According to these results, the EtOAc-F from G. ulmifolia leaves exhibit anthelmintic effect and increased the stress biomarkers on H. contortus.

Stress, Allostatic Load, and Neuroinflammation: Implications for Racial and Socioeconomic Health Disparities in Glaucoma.

Glaucoma is the leading cause of irreversible blindness, and its pathophysiology includes neuroinflammatory changes. The present therapies for glaucoma target pressure-lowering mechanisms with limited success, making neuroinflammation a target for future interventions. This review summarizes the neuroinflammatory pathways seen in glaucoma and their interplay with stress. Glucocorticoids have been shown to activate proinflammatory glial cells, contributing to the neuroinflammation in glaucoma. Glucocorticoids have also been shown to increase the IOP directly. Stress-associated autonomic dysfunction can affect the vascular homeostasis in the retina and create oxidative stress. Diabetes, hyperglycemic-mediated endothelial damage, and vascular inflammation also play important roles in the neuroinflammation in glaucoma and diabetic retinopathy. Psychosocial stress has been implicated in an increased IOP and glaucoma outcomes. People who experience maladaptive chronic stress suffer from a condition known as allostatic load, which describes pathologic neuroendocrine dysregulation. The effects of allostatic load and chronic stress have been studied in patients affected by a lower socioeconomic status (SES) and marginalized racial identities. A lower SES is associated with higher rates of glaucoma and also affects the access to care and screening. Additionally, people of African ancestry are disproportionately affected by glaucoma for reasons that are multifactorial. In conclusion, this review explores neuroinflammation in glaucoma, highlighting opportunities for future investigation.

Bartonella spp. in different species of bats from Misiones (Argentina).

The aim of this study was to detect vector-borne pathogens (Anaplasmataceae family, Rickettsia genus, and Bartonella genus) in bats from Misiones (Argentina). Thirty-three specimens were captured over 8 days using mist nets. Twenty (60.6%) blood samples were positive (11/13 Artibeus lituratus, 4/10 Desmodus rotundus, 4/8 Carollia perspicillata, and 1/2 Myotis nigricans) by PCR for the gltA gene fragment of Bartonella. All samples were negative by PCR for the Anaplasmataceae family and Rickettsia genus. The phylogenetic analysis showed seven Bartonella genotypes. The three genotypes obtained from A. lituratus, 2 from C. perspicillata, and 1 from D. rotundus were related to Bartonella spp. from New World bats, while the sequence obtained from M. nigricans was related to Old World bats. We identified a considerable diversity of Bartonella genotypes in a small number of bats, thus further research is required to better understand the complex bat-pathogen interaction.

Interferon γ, IL-17, and IL-1ß impair sperm motility and viability and induce sperm apoptosis.

Urogenital inflammation is a known cause of male infertility. Increased levels of inflammatory cytokines, leukocyte counts and oxidative stress are highly detrimental for sperm quality thus compromising male fertility. Although cytokines affect sperm by recruiting and activating leukocytes consequently inducing tissue inflammation and oxidative stress, scarce to absent data have been reported about the putative direct effects of inflammatory cytokines on spermatozoa. Herein, we analyzed whether IFNγ, IL-17A, IL-1ß, and IL-8 can alter human sperm motility and viability per se. Fractions of viable and motile spermatozoa from normospermic healthy donors were in vitro incubated with recombinant human IFNγ, IL-17A, IL-1ß or IL-8 and sperm ROS production, motility, viability and apoptosis were analyzed. Sperm exposed to different concentrations of IFNγ, IL-17A and IL-1ß, or a combination of them, for either 1 or 3 h showed significantly increased levels of mitochondrial ROS production and reduced motility and viability with respect to sperm incubated with vehicle. Moreover, the exposure to IFNγ, IL-17A and IL-1ß resulted in significantly higher levels of early and/or late apoptotic and/or necrotic spermatozoa. Interestingly, no significant differences in sperm motility, viability and apoptosis were observed in sperm incubated with the concentrations of IL-8 analyzed, for either 1 or 3 h, with respect to sperm incubated with vehicle. In conclusion, our results indicate that IFNγ, IL-17A and IL-1ß per se impair sperm motility and decreases viability by triggering increased mitochondrial ROS production and inducing sperm apoptosis. Our results suggest that screening inflammatory cytokines in semen would be an additional helpful tool for the diagnostic workup of male infertility.

A Conceptual Framework to Integrate Biodiversity, Ecosystem Function, and Ecosystem Service Models.

Global biodiversity and ecosystem service models typically operate independently. Ecosystem service projections may therefore be overly optimistic because they do not always account for the role of biodiversity in maintaining ecological functions. We review models used in recent global model intercomparison projects and develop a novel model integration framework to more fully account for the role of biodiversity in ecosystem function, a key gap for linking biodiversity changes to ecosystem services. We propose two integration pathways. The first uses empirical data on biodiversity-ecosystem function relationships to bridge biodiversity and ecosystem function models and could currently be implemented globally for systems and taxa with sufficient data. We also propose a trait-based approach involving greater incorporation of biodiversity into ecosystem function models. Pursuing both approaches will provide greater insight into biodiversity and ecosystem services projections. Integrating biodiversity, ecosystem function, and ecosystem service modeling will enhance policy development to meet global sustainability goals.

Phytochemical Compounds and Pharmacological Properties of Larrea tridentata.

For centuries, traditional medicine from plants (phytotherapy) was the only treatment for infectious and non-infectious diseases. Although it is still practiced in several countries with excellent therapeutic results, it is frequently underestimated because, unlike Western medicine, it is not based on an empirical scientific foundation. However, interest in the search for plant-based therapeutic resources has been stimulated by disciplines such as phytochemistry and the side effects of conventional pharmacological therapies. For example, Larrea tridentata is a perennial shrub used in traditional medicine in northern Mexico and the southern United States to treat infertility, rheumatism, arthritis, colds, diarrhea, skin problems, pain, inflammation and excess body weight. Scientific research has revealed its beneficial effects-antioxidant, antitumor, neuroprotective, regenerative, antibacterial, antiviral, antifungal, anthelmintic, antiprotozoal and insecticidal-although reports indicate that some compounds in Larrea tridentata may be hepatotoxic and nephrotoxic. Therefore, the aim of this review was to highlight the updates regarding phytochemical compounds and the pharmacological properties of Larrea tridentata.

An Artemisia cina n-hexane extract reduces the Haemonchus contortus and Teladorsagia circumcincta fecal egg count in naturally infected periparturient goats.

The objective of this study was to evaluate an n-hexane extract of Artemisia cina (Acn-h) as a natural anthelmintic treatment for periparturient goats naturally infected with the nematodes Haemonchus contortus and Teladorsagia circumcincta. A total of 200 periparturient Alpine and Nubian goats were used. Deworming criteria were based on the following parameters: fecal egg account (epg), ocular mucosa color (OMC), and body condition (BC). A previous analysis using coprocultures of the flock revealed the presence of H. contortus (80%) and T. circumcincta (20%). The Acn-h contained two new compounds identified by mass spectrometry data as isoguaiacin and norisoguaiacin at 284.14 and 315 m/z. The lethal effects of Acn-h at 0.5, 1, 2, and 4 mg/mL on H. contortus adult stages were 31.6, 66.5, 81.3, and 86.9%, respectively (p < 0.05), showing similar efficacy at 2 and 4 mg/mL with albendazole (positive control group). Then, two experimental groups, with 100 goats in peripartum in each, were distributed randomly and treated at day 0 as follows: group 1 = 4 mg/kg of Acn-h as single oral dose, and group 2 = control group, treated with water (as a placebo). The epg, OMC, and BC parameters were recorded at 0 (periparturient period), 7 (birth period), and 23 (postpartum) days and analyzed using a completely randomized design with Duncan's test for comparison of means and analysis of variance. The following epg reductions were recorded in the Acn-h-treated group as follows: 20.1 ± 34.4 and 31.7 ± 38.2% at days 7 and 23 compared to the control group. During the whole experiment, no significant differences in OMC or BC were observed in relation to the control group, excepting at day 23 (p < 0.05) for BC in the group treated with A. cina. Thus, Acn-h can be a useful natural alternative tool for the control of the nematodes H. contortus and T. circumcincta in periparturient goat flocks.

Medicago truncatula Yellow Stripe-Like7 encodes a peptide transporter participating in symbiotic nitrogen fixation.

Yellow Stripe-Like (YSL) proteins are a family of plant transporters that are typically involved in transition metal homeostasis. Three of the four YSL clades (I, II and IV) transport metals complexed with the non-proteinogenic amino acid nicotianamine or its derivatives. No such capability has been shown for any member of clade III, but the link between these YSLs and metal homeostasis could be masked by functional redundancy. We studied the role of the clade III YSL protein MtSYL7 in Medicago truncatula nodules. MtYSL7, which encodes a plasma membrane-bound protein, is mainly expressed in the pericycle and cortex cells of the root nodules. Yeast complementation assays revealed that MtSYL7 can transport short peptides. M. truncatula transposon insertion mutants with decreased expression of MtYSL7 had lower nitrogen fixation rates and showed reduced plant growth whether grown in symbiosis with rhizobia or not. YSL7 mutants accumulated more copper and iron in the nodules, which is likely to result from the increased expression of iron uptake and delivery genes in roots. Taken together, these data suggest that MtYSL7 plays an important role in the transition metal homeostasis of nodules and symbiotic nitrogen fixation.

Association between meteorological variables and semen quality: a retrospective study.

Spermatogenesis is a temperature-dependent process, and high summer temperatures have been linked to lower sperm concentration and count. However, reports describing the association between other meteorological variables and semen quality are scarce. This study evaluated the association between semen quality and temperature, humidity, pressure, apparent temperature (AT), temperature-humidity index (THI), simplified wet-bulb global temperature (sWBGT), and sunshine duration. Semen samples were obtained at the Laboratorio de Andrología y Reproducción (LAR, Argentina), from men undergoing routine andrology examination (n=11657) and computer-assisted sperm analysis (n=4705) following WHO 2010 criteria. Meteorological variables readings were obtained from the Sistema Meteorológico Nacional. Sperm quality parameters were negatively affected in summer when compared to winter. Additionally, there was a significant decrease in sperm kinematics between winter and spring. Branch and bound variable selection followed by multiple regression analysis revealed a significant association between semen quality and meteorological variables. Specifically, changes in sunshine duration and humidity reinforced the prognosis of semen quality. Highest/lowest sunshine duration and humidity quantiles resulted in decreased sperm concentration, count, motility, vitality and membrane competence, nuclear maturity, and sperm kinematics associated to highest sunshine duration and lowest humidity. Findings from this report highlight the relevance of environmental studies for predicting alterations in male reproductive health associated to variations in meteorological variables, especially considering the current climate changes around the planet due to global warming and its consequences for human health.

Reproductive biology of the swell shark Cephaloscyllium ventriosum (Carcharhiniformes: Scyliorhinidae).

The Cephaloscyllium ventriosum shark is present in the artisanal fisheries of elasmobranchs on the western coast of Baja California Sur, Mexico. The main characteristics of the sexual maturation of this species based on individuals captured from off north-west Mexico in 2013-2016 are described. The size at maturity of this species was determined for the first time (total length 82 cm for females and 76 cm for males). Most females had one egg case per one uterus, and two per one uterus was an isolated event of low incidence. From the histological analysis of females, it was possible to show sperm storage in the oviducal gland. Fully developed sperm in immature organisms were identified in the testes. The main indicator of the maturity stage of males and their mating activity is the clasper. The present study provides evidence for a reliable estimation of the sexual maturity of these organisms, demonstrating the need for the combination of macroscopic and microscopic methods.

Reproductive biology of the Rasptail skate Rostroraja velezi (Rajiformes: Rajidae).

The Rasptail skate Rostroraja velezi is commercially exploited in artisanal elasmobranch fisheries along the west coast of Baja California Sur, Mexico, but information on its life history is limited. This study aimed to investigate the reproductive biology of R. velezi. A total of 105 specimens were caught from April 2008 to May 2012, including the largest reported specimen with 121 cm total length, 96 cm disc width (DW ). Females attained larger sizes than males. Males and females presented functional gonads. There was an asymmetry in the testes of males, with the left testis being larger. Histological analysis of the reproductive biology of R. velezi was performed here for the first time. The presence of sperm storage in females and spermatogenic development beginning at the first stages of maturity in males was recorded. It was possible to identify the development of secretions in the club, baffle and terminal zone of the oviducal gland. DW at maturity, defined as the DW at which 50% of the population is mature, was estimated at 68-72 cm for females and 65.1 cm for males. Egg-bearing females caught in April and May presented one egg capsule per uterus. Furthermore, a description of the egg capsule of R. velezi is provided. Elucidating the reproductive cycle, the type of reproductive strategies, and the fecundity of R. velezi will allow us to understand the impact of fisheries on this species.

Medicago truncatula Ferroportin2 mediates iron import into nodule symbiosomes.

Iron is an essential cofactor for symbiotic nitrogen fixation, required by many of the enzymes involved, including signal transduction proteins, O2 homeostasis systems, and nitrogenase itself. Consequently, host plants have developed a transport network to deliver essential iron to nitrogen-fixing nodule cells. Ferroportin family members in model legume Medicago truncatula were identified and their expression was determined. Yeast complementation assays, immunolocalization, characterization of a tnt1 insertional mutant line, and synchrotron-based X-ray fluorescence assays were carried out in the nodule-specific M. truncatula ferroportin Medicago truncatula nodule-specific gene Ferroportin2 (MtFPN2) is an iron-efflux protein. MtFPN2 is located in intracellular membranes in the nodule vasculature and in inner nodule tissues, as well as in the symbiosome membranes in the interzone and early-fixation zone of the nodules. Loss-of-function of MtFPN2 alters iron distribution and speciation in nodules, reducing nitrogenase activity and biomass production. Using promoters with different tissular activity to drive MtFPN2 expression in MtFPN2 mutants, we determined that expression in the inner nodule tissues is sufficient to restore the phenotype, while confining MtFPN2 expression to the vasculature did not improve the mutant phenotype. These data indicate that MtFPN2 plays a primary role in iron delivery to nitrogen-fixing bacteroids in M. truncatula nodules.

The importance of Legal Reserves for protecting the Pantanal biome and preventing agricultural losses.

Considering scenarios of future changes in land use have the potential to support policy-makers in drafting environmental laws to reconcile the demands of multiple land uses. The Pantanal, one of the largest wetlands in the world, has been undergoing rapid land use changes, and does not yet have any integrated environmental legislation on Legal Reserve for entire region (LR - minimum percentage of native vegetation required within private properties). The aim of this paper was to generate future vegetation loss scenarios for the Pantanal based on four LR values: (i) BAU: Business as usual, which considers existing laws: Native Vegetation Protection Law and State Decree; (ii) LRE: LR elimination owing to a bill recently proposed; (iii) LR50: which considers the bill proposing 50% of LR for the Pantanal; and (iv) LR80: our proposed levels of 80% of LR for the lowlands and 35% for the plateau (following values in the Amazon). Based on native vegetation loss from each scenario, we estimated the soil loss and sediment yield to rivers. Our results show that LRE would increase native vegetation loss in the Pantanal by as much as 139% when compared to the BAU, whereas increasing LR levels would reduce conversion values by 29% (LR80). Elimination of the LR would increase soil erosion and sediment production by up to 7% and 10%, respectively, compared to BAU. Based on native vegetation loss from each scenario, we estimated the soil loss and sediment yield to rivers with our data showing more than 90% of the sediment transported to the lowland originating from the plateau. The LR80 indicates a reduction in soil nutrient replacement costs of 10% compared to BAU, while in the LR50 these costs decrease by 1.5%, and in the LRE would increase of 8%. Our results show that abolishing current protections would have substantial impacts on avulsion processes, on several economic activities (tourism, fishery, cattle raising, etc.) and negative impacts for biodiversity conservation and would bring losses to agriculture in the Pantanal. Hence, our study brings clearly evidence of LR importance and need to expand it in this sensitive wetland.

Peroral endoscopic myotomy in pediatric jackhammer esophagus.

The jackhammer esophagus is a rare hypercontractile disorder and diagnosis is based on high-resolution manometry. Peroral endoscopic myotomy (POEM) of the spastic esophagus segments has been described. We report a pediatric patient with jackhammer esophagus that was treated endoscopically.

Spatial and temporal dimensions of landscape fragmentation across the Brazilian Amazon.

The Brazilian Amazon in the past decades has been suffering severe landscape alteration, mainly due to anthropogenic activities, such as road building and land clearing for agriculture. Using a high-resolution time series of land cover maps (classified as mature forest, non-forest, secondary forest) spanning from 1984 through 2011, and four uncorrelated fragmentation metrics (edge density, clumpiness index, area-weighted mean patch size and shape index), we examined the temporal and spatial dynamics of forest fragmentation in three study areas across the Brazilian Amazon (Manaus, Santarém and Machadinho d'Oeste), inside and outside conservation units. Moreover, we compared the impacts on the landscape of: (1) different land uses (e.g. cattle ranching, crop production), (2) occupation processes (spontaneous vs. planned settlements) and (3) implementation of conservation units. By 2010/2011, municipalities located along the Arc of Deforestation had more than 55% of the remaining mature forest strictly confined to conservation units. Further, the planned settlement showed a higher rate of forest loss, a more persistent increase in deforested areas and a higher relative incidence of deforestation inside conservation units. Distinct agricultural activities did not lead to significantly different landscape structures; the accessibility of the municipality showed greater influence in the degree of degradation of the landscapes. Even with a high proportion of the landscapes covered by conservation units, which showed a strong inhibitory effect on forest fragmentation, we show that dynamic agriculturally driven economic activities, in municipalities with extensive road development, led to more regularly shaped, heavily fragmented landscapes, with higher densities of forest edge.

Thresholds of species loss in Amazonian deforestation frontier landscapes.

In the Brazilian Amazon, private land accounts for the majority of remaining native vegetation. Understanding how land-use change affects the composition and distribution of biodiversity in farmlands is critical for improving conservation strategies in the face of rapid agricultural expansion. Working across an area exceeding 3 million ha in the southwestern state of Rondônia, we assessed how the extent and configuration of remnant forest in replicate 10,000-ha landscapes has affected the occurrence of a suite of Amazonian mammals and birds. In each of 31 landscapes, we used field sampling and semistructured interviews with landowners to determine the presence of 28 large and medium sized mammals and birds, as well as a further 7 understory birds. We then combined results of field surveys and interviews with a probabilistic model of deforestation. We found strong evidence for a threshold response of sampled biodiversity to landscape level forest cover; landscapes with <30-40% forest cover hosted markedly fewer species. Results from field surveys and interviews yielded similar thresholds. These results imply that in partially deforested landscapes many species are susceptible to extirpation following relatively small additional reductions in forest area. In the model of deforestation by 2030 the number of 10,000-ha landscapes under a conservative threshold of 43% forest cover almost doubled, such that only 22% of landscapes would likely to be able to sustain at least 75% of the 35 focal species we sampled. Brazilian law requires rural property owners in the Amazon to retain 80% forest cover, although this is rarely achieved. Prioritizing efforts to ensure that entire landscapes, rather than individual farms, retain at least 50% forest cover may help safeguard native biodiversity in private forest reserves in the Amazon.

Modelling land cover change in the Brazilian Amazon: temporal changes in drivers and calibration issues.

Land cover change (LCC) models are used in many studies of human impacts on the environment, but knowing how well these models predict observed changes in the landscape is a challenge. We used nearly three decades of LCC maps to run several LCC simulations to: (1) determine which parameters associated with drivers of LCC (e.g. roads) get selected for which transition (forest to deforested, regeneration to deforested or deforested to regeneration); (2) investigate how the parameter values vary through time with respect to the different activities (e.g. farming); and (3) quantify the influence of choosing a particular time period for model calibration and validation on the performance of LCC models. We found that deforestation of primary forests tends to occur along roads (included in 95 % of models) and outside protected areas (included in all models), reflecting farming establishment. Regeneration tends to occur far from roads (included in 78 % of the models) and inside protected areas (included in 38 % of the models), reflecting the processes of land abandonment. Our temporal analysis of model parameters revealed a degree of variation through time (e.g. effectiveness of protected areas rose by 73 %, p < 0.001), but for the majority of parameters there was no significant trend. The degree to which model predictions agreed with observed change was heavily dependent on the year used for calibration (p < 0.001). The next generation of LCC models may need to embed trends in parameter values to allow the processes determining LCC to change through time and exert their influence on model predictions.