Spatiotemporal Imaging of Zinc Ions in Zebrafish Live Brain Tissue Enabled by Fluorescent Bionanoprobes

The zebrafish is a powerful model organism to study the mechanisms governing transition metal ions within whole brain tissue. Zinc is one of the most abundant metal ions in the brain, playing a critical pathophysiological role in neurodegenerative diseases. The homeostasis of free, ionic zinc (Zn2+) is a key intersection point in many of these diseases, including Alzheimer’s disease and Parkinson’s disease. A Zn2+ imbalance can eventuate several disturbances that may lead to the development of neurodegenerative changes. Therefore, compact, reliable approaches that allow the optical detection of Zn2+ across the whole brain would contribute to our current understanding of the mechanisms that underlie neurological disease pathology. We developed an engineered fluorescence protein-based nanoprobe that can spatially and temporally resolve Zn2+ in living zebrafish brain tissue. The self-assembled engineered fluorescence protein on gold nanoparticles was shown to be confined to defined locations within the brain tissue, enabling site specific studies, compared to fluorescent protein-based molecular tools, which diffuse throughout the brain tissue. Two-photon excitation microscopy confirmed the physical and photometrical stability of these nanoprobes in living zebrafish (Danio rerio) brain tissue, while the addition of Zn2+ quenched the nanoprobe fluorescence. Combining orthogonal sensing methods with our engineered nanoprobes will enable the study of imbalances in homeostatic Zn2+ regulation. The proposed bionanoprobe system offers a versatile platform to couple metal ion specific linkers and contribute to the understanding of neurological diseases

Upper ocean temperature characteristics in the subantarctic Southeast Pacific based on biomarker reconstructions

Alkenones and Glycerol Dialkyl Glycerol Tetraether lipids (GDGT) as remnants of living organisms are widely used biomarkers for determining past oceans&rsquo; water temperatures. The organisms these proxy carriers stem from, are influenced by a number of environmental parameters, such as water depth, nutrient availability, light conditions or seasonality, which all may significantly bias the calibration to ambient water temperatures. Reliable temperature determinations remain thus challenging, especially in higher latitudes and for under-sampled regions. We analyzed 33 sediment surface samples from the Southern Chilean continental margin and the Drake Passage for alkenones and GDGTs and compared the results with gridded instrumental reference data from the World Ocean Atlas 2005 (WOA05), as well as previously published data from an extended study area covering the Central and Western South Pacific towards the New Zealand continental margin. We show that for alkenone-derived SSTs, the widely-used global core-top calibration of M&uuml;ller et al. (1998) yields the smallest residuals, whereas the calibration of Sikes et al. (1997), adapted to higher latitudes and supposed to show summer SSTs, overestimates modern WOA05-based (summer and annual mean) SSTs. Our alkenone SSTs show a slight seasonal shift of ~1 &deg;C at the Southern Chilean Margin and up to ~2 &deg;C in the Drake Passage towards austral summer SSTs, whereas samples in the Central South Pacific reflect an annual mean signal. We show that for GDGT-based temperatures, a more complex pattern emerges. In areas north of the Subantarctic Front (SAF) the subsurface calibration of Kim et al. (2012a) best reflects temperatures from the WOA05, largely within the margin error of &plusmn;2.2 &deg;C. Temperatures south of the SAF instead are significantly overestimated by up to 14 &deg;C, irrespective of the applied calibration. Based on a qualitative assessment of the GDGT [2] / [3]-ratios, which likely indicate water depth of origin, our samples reflect a subsurface (0 to 200 m water depth) rather than a surface (0&ndash;50 m water depth) signal. The overestimation of surface and subsurface temperatures south of the SAF highlights the need for a re-assessment of existing calibrations in the polar Southern Ocean, and leads to limitations in reliably both obtaining absolute values and assessing relative changes. Therefore, we suggest a modified Southern Ocean calibration for surface and subsurface GDGT-based temperatures, which shows a lower temperature sensitivity of the TEXL86 and yields principally lower absolute temperatures, which align more closely with WOA05-derived values.</p

Non-intersecting squared Bessel paths and multiple orthogonal polynomials for modified Bessel weights

We study a model of $n$ non-intersecting squared Bessel processes in the confluent case: all paths start at time $t = 0$ at the same positive value $x = a$, remain positive, and are conditioned to end at time $t = T$ at $x = 0$. In the limit $n \to \infty$, after appropriate rescaling, the paths fill out a region in the $tx$-plane that we describe explicitly. In particular, the paths initially stay away from the hard edge at $x = 0$, but at a certain critical time $t^*$ the smallest paths hit the hard edge and from then on are stuck to it. For $t \neq t^*$ we obtain the usual scaling limits from random matrix theory, namely the sine, Airy, and Bessel kernels. A key fact is that the positions of the paths at any time $t$ constitute a multiple orthogonal polynomial ensemble, corresponding to a system of two modified Bessel-type weights. As a consequence, there is a $3 \times 3$ matrix valued Riemann-Hilbert problem characterizing this model, that we analyze in the large $n$ limit using the Deift-Zhou steepest descent method. There are some novel ingredients in the Riemann-Hilbert analysis that are of independent interest.Comment: 59 pages, 11 figure

Appraising and applying evidence about a diagnostic test during a performance-based assessment

BACKGROUND: The practice of Evidence-based Medicine requires that clinicians assess the validity of published research and then apply the results to patient care. We wanted to assess whether our soon-to-graduate medical students could appraise and apply research about a diagnostic test within a clinical context and to compare our students with peers trained at other institutions. METHODS: 4(th )year medical students who previously had demonstrated competency at probability revision and just starting first-year Internal Medicine residents were used for this research. Following an encounter with a simulated patient, subjects critically appraised a paper about an applicable diagnostic test and revised the patient's pretest probability given the test result. RESULTS: The medical students and residents demonstrated similar skills at critical appraisal, correctly answering 4.7 and 4.9, respectively, of 6 questions (p = 0.67). Only one out of 28 (3%) medical students and none of the 15 residents were able to correctly complete the probability revision task (p = 1.00). CONCLUSIONS: This study found that most students completing medical school are able to appraise an article about a diagnostic test but few are able to apply the information from the article to a patient. These findings raise questions about the clinical usefulness of the EBM skills possessed by graduating medical students within the area of diagnostic testing