Realistic levels of a fertilizer impair Iberian newt embryonic development

A wide variety of agricultural chemicals with potential to affect amphibian health are released into the environment daily. Most of these chemicals are xenobiotic compounds that are highly toxic to embryos, tadpoles, and terrestrial stages. Other substances that occur in pristine environments at harmless concentrations, such as inorganic nitrogenous compounds, may reach potentially toxic levels as a consequence of certain human activities, including the application of fertilizers. Most of the studies that analyze the effects of inorganic nitrogen on amphibian embryos are conducted with anurans, whereas little information exists regarding urodeles. Embryos of newts and salamanders usually exhibit longer times to hatch than frogs and toads. A longer hatching time results in a longer exposure of embryos to diffuse environmental pollution and therefore a higher risk of suffering toxic effects during the embryonic stage. We demonstrate that ammonium nitrate, a widely used nitrogenous fertilizer, at concentrations used in areas of high agricultural intensity, affects embryonic development of the Iberian newt (Lissotriton boscai). Although ammonium nitrate did not have significant lethal effects, it reduced the growth rate of exposed embryos, which were 9.6% smaller than controls. Hatching synchrony remained similar across treatments, and hatching date was not affected by ammonium nitrate, indicating that the effect on growth was not time- dependent. Researchers have demonstrated fitness costs in smaller than average tadpoles, suggesting that ammonium nitrate exposure could have long-term negative consequences for the Iberian newt.Peer reviewe

Adaptation to osmotic stress provides protection against ammonium nitrate in Pelophylax perezi embryos

The negative effects of pollution on amphibians are especially high when animals are additionally stressed by other environmental factors such as water salinity. However, the stress provoked by salinity may vary among populations because of adaptation processes. We tested the combined effect of a common fertilizer, ammonium nitrate (0–90.3 mg N–NO3NH4/L), and water salinity (0–2‰) on embryos of two Pelophylax perezi populations from ponds with different salinity concentrations. Embryos exposed to the fertilizer were up to 17% smaller than controls. Survival rates of embryos exposed to a single stressor were always below 10%. The exposure to both stressors concurrently increased mortality rate (>95%) of embryos from freshwater. Since the fertilizer was lethal only when individuals were stressed by the salinity, it did not cause lethal effects on embryos naturally adapted to saline environments. Our results underscore the importance of testing multiple stressors when analyzing amphibian sensitivity to environmental pollution.Funding was provided by the Regional Government of Castilla y León (SA071A06) and the Ministry of Science and Innovation of Spain (CGL2005-0372).Peer reviewe

Influence of a combination of agricultural chemicals on embryos of the endangered gold-striped salamander (Chioglossa lusitanica)

Pollution from agrochemicals may be contributing to the global decline of amphibian populations. Environmentally relevant concentrations of a fertiliser, ammonium nitrate, and a commercial formulation of the herbicide glyphosate Roundup Plus were tested on the embryonic development of Chioglossa lusitanica. This study introduces new data at three different levels. First, we provide previously unknown information about hatchling traits of C. lusitanica. Second, we present the first ecotoxicological study of this endangered species, to which environmental pollution is considered a major threat. Third, we conduct the first experiment with an amphibian species exposed to a mixture of a glyphosate-based herbicide and a nitrogenous fertiliser. Control individuals hatched with an average (±SD) total length of 18.77 (±2.02) mm and at an average Harrison’s developmental stage of 44.58 (±1.24). Mean hatching time among controls was 11.52 (±1.29) weeks. None of the chemicals or their interaction produced lethal effects; however, a significant interaction was found when analysing total length at hatching. Individuals exposed to the herbicide hatched at a larger size than controls, and this effect was especially clear when the fertiliser was added to the water. The absence of pollutant-related mortality or severe sublethal effects is in agreement with most studies indicating a high tolerance of amphibian embryos to agrochemicals. However, further research considering other life stages and additional natural factors (i.e., predators, food availability) is needed to estimate the ecological impact of chemical mixtures on C. lusitanica.Peer reviewe

Selective isolation of mouse glial nuclei optimized for reliable downstream omics analyses

Background: Isolation of cell types of interest from the brain for molecular applications presents several challenges, including cellular damage during tissue dissociation or enrichment procedures, and low cell number in the tissue in some cases. Techniques have been developed to enrich distinct cell populations using immunopanning or fluorescence activated cell/nuclei sorting. However, these techniques often involve fixation, immunolabeling and DNA staining steps, which could potentially influence downstream omics applications. New method: Taking advantage of readily available genetically modified mice with fluorescent-tagged nuclei, we describe a technique for the purification of cell-type specific brain nuclei, optimized to decrease sample preparation time and to limit potential artefacts for downstream omics applications. We demonstrate the applicability of this approach for the purification of glial cell nuclei and show that the resulting cell-type specific nuclei obtained can be used effectively for omics applications, including ATAC-seq and RNA-seq. Results: We demonstrate excellent enrichment of fluorescently-tagged glial nuclei, yielding high quality RNA and chromatin. We identify several critical steps during nuclei isolation that help limit nuclei rupture and clumping, including quick homogenization, dilution before filtration and loosening of the pellet before resuspension, thus improving yield. Sorting of fluorescent nuclei can be achieved without fixation, antibody labelling, or DAPI staining, reducing potential artifactual results in RNA-seq and ATAC-seq analyses. We show that reproducible glial cell type-specific profiles can be obtained in transcriptomic and chromatin accessibility assays using this rapid protocol. Comparison with existing methods: Our method allows for rapid enrichment of glial nuclei populations from the mouse brain with minimal processing steps, while still providing high quality RNA and chromatin required for reliable omics analyses. Conclusions: We provide a reproducible method to obtain nucleic material from glial cells in the mouse brain with a quick and limited sample preparation

Diagnóstico matemático de ecocardiografías pediátricas con medidas de dimensión fractal evaluadas con armonía matemática intrínseca

Background and objectivesGeometry allows the objective mathematical characterization of forms. Fractal geometry characterizes irregular objects. The left ventricle dynamical states form observed through echocardiography can be objectively evaluated through fractal dimension measures.MethodsA measurement of fractal dimension was performed using the Box-counting method of three defined objects in 28 echocardiographic images, 16 from normal children (group A) and 12 ill children (group B), in order to establish differences between health and illness from its comparison with the fractal dimensions of 2 normality prototypes and 2 disease prototypes.ResultsA new diagnostic, clinical application methodology was developed based in the “intrinsic mathematical harmony“(IMH) concept, and it was observed that the fractal dimensions of the defined objects for an abnormal echocardiogram show similarity to its fourth significant number, thus demonstrating the possibility of following up the evolution from normality towards disease. According to the performed calculations, 68.75% of the cases in group A could be better evaluated with the developed diagnostic methodology, and the ill ones could be diagnosed more effectively.ConclusionsThe pediatric echocardiography images can be objectively characterized with fractal dimension measurements, thus enabling the development of a clinical diagnostic methodology of echocardiography in children from the IMH concep

Physical States in Canonically Quantized Supergravity

We discuss the canonical quantization of $N=1$ supergravity in the functional Schrodinger representation. Although the form of the supersymmetry constraints suggests that there are solutions of definite order $n$ in the fermion fields, we show that there are no such states for any finite $n$. For $n=0$, a simple scaling argument definitively excludes the purely bosonic states discussed by D'Eath. For $n>0$, the argument is based on a mode expansion of the gravitino field on the quantization 3-surface. It is thus suggested that physical states in supergravity have infinite Grassmann number. This is confirmed for the free spin-3/2 field, for which we find that states satisfying the gauge constraints contain an infinite product of fermion mode operators.Comment: 36 pages (uses jnl.tex), CTP #227

Spin/disorder correlations and duality in the c=1/2 string

We use the method of discrete loop equations to calculate exact correlation functions of spin and disorder operators on the sphere and on the boundary of a disk in the $c = 1/2$ string, both in the Ising and dual Ising matrix model formulations. For both the Ising and dual Ising theories the results on the sphere are in agreement with the KPZ/DDK scaling predictions based on Liouville theory; the results on the disk agree with the scaling predictions of Martinec, Moore, and Seiberg for boundary operators. The calculation of Ising disorder correlations on the sphere requires the use of boundary variables introduced in [hep-th/9510199], which have no matrix model analog. A subtlety in the calculation on the disk arises because the expansions of the correlation functions have leading singular terms which are nonuniversal; we show that this issue may be resolved by using separate cosmological constants for each boundary domain. These results give evidence that the Kramers-Wannier duality symmetry of the $c = 1/2$ conformal field theory survives coupling to quantum gravity, implying a duality symmetry of the $c = 1/2$ string even in the presence of boundary operators.Comment: 19 pages, LaTeX; final publication versio

Diagnosis of multiple sclerosis using optical coherence tomography supported by artificial intelligence

Background: Current procedures for diagnosing multiple sclerosis (MS) present a series of limitations, making it critically important to identify new biomarkers. The aim of the study was to identify new biomarkers for the early diagnosis of MS using spectral-domain optical coherence tomography (OCT) and artificial intelligence. Methods: Spectral domain OCT was performed on 79 patients with relapsing-remitting multiple sclerosis (RRMS) (disease duration ≤ 2 years, no history of optic neuritis) and on 69 age-matched healthy controls using the posterior pole protocol that incorporates the anatomic Positioning System. Median retinal thickness values in both eyes and inter-eye difference in healthy controls and patients were evaluated by area under the receiver operating characteristic (AUROC) curve analysis in the foveal, parafoveal and perifoveal areas and in the overall area spanned by the three rings. The structures with the greatest discriminant capacity — retinal thickness and inter-eye difference — were used as inputs to a convolutional neural network to assess the diagnostic capability. Results: Analysis of retinal thickness and inter-eye difference in RRMS patients revealed that greatest alteration occurred in the ganglion cell (GCL), inner plexiform (IPL), and inner retinal (IRL) layers. By using the average thickness of the GCL (AUROC = 0.82) and the inter-eye difference in the IPL (AUROC = 0.71) as inputs to a two-layer convolutional neural network, automatic diagnosis attained accuracy = 0.87, sensitivity = 0.82, and specificity = 0.92. Conclusion: This study adds weight to the argument that neuroretinal structure analysis could be incorporated into the diagnostic criteria for MS