Reading projects

"By reading only six hours a day", says Marianne Dashwood, outlining her plan of future application to her sister Elinor in Sense and Sensibility, "I shall gain in the course of a twelve-month a great deal of instruction which I now feel myself to want." She adds: "Our own library is too well known to me, to be resorted to for any thing beyond mere amusement. But there are many works well worth reading at the Park; and there are others of more modern production which I know I can borrow of Colonel Brandon" (301). We know, to some extent, what was in the Dashwoods' own library – volumes of Cowper, Scott and Thomson are mentioned. But what might Marianne have borrowed at Barton Park and Delaford? Which publications would Colonel Brandon have considered most appropriate for her project of self-improvement? Elinor considers Marianne's plan excessive, but what would have been a more realistic amount of time for her to spend reading each day, and where might she have done it

Additional file 1: of PIGO deficiency: palmoplantar keratoderma and novel mutations

Gene selection from WES data. (DOCX 105 kb

Phenotype analysis, nephrin expression and evaluation of glomerular function in adriamycin exposed zebrafish.

<p>(A) Different categories of phenotype were defined in the adriamycin exposed embryos at 4 dpf: embryos with a normal phenotype, embryos with visible pericardial edema (white arrow) and dysmorphic or dead embryos. (B) 100 embryos per condition were live-screened at 4 dpf and assigned to a phenotype category. Increasing adriamycin concentrations in the culture medium were associated with an increasing percentage of embryos with pericardial edema. (C) qPCR was performed using total RNA from control and adriamycin exposed embryos at 4 dpf. A significantly decreased expression of nephrin (corrected for housekeeping gene <i>elfa</i>) was observed in the adriamycin exposed embryos compared to the control fish. The experiment was performed twice in triplicate. Bars represent means ± SD. * P<0.05 in comparison to condition without the addition of adriamycin. (D) Rhodamine-labeled 70 kDa dextran was injected in the cardiac venous sinus of 75 hpf old embryos. LEFT: A representative immunofluorescence picture of a control embryo immediately after injection shows the distribution of fluorescence through the vascular system of the zebrafish larva. A dose-dependent diminishing effect of adriamycin on fluorescence recorded in the fish eye 5 hours after injection was observed. Representative images of the eye from 0, 10 and 30 μM adriamycin treated embryos 5 hours after injection are shown. RIGHT: A diagrammatic representation shows the quantification of the mean fluorescence intensity ± SD recorded in the retinal vascular bed. * P < 0.05 in comparison to condition without the addition of adriamycin.</p

Additional file 1: Table S1. of Platelet studies in autism spectrum disorder patients and first-degree relatives

Criteria leading to exclusion of individuals in the platelet study. (DOC 33 kb

Additional file 2: Figure S1. of Platelet studies in autism spectrum disorder patients and first-degree relatives

Platelet activation pathways investigated in this study. Platelet aggregation with the strong agonist arachidonic acid (AA) stimulates platelets via the formation of thromboxane A2 (TxA2) by the enzyme COX1 to finally active the G protein-coupled thromboxane receptor that will result in calcium efflux and the release of alpha (α) and dense (δ) granules. A high dose of AA does not require platelet secretion for full platelet activation. Activation with the weak agonist epinephrine (EPI) will partially stimulate platelets via the adrenergic receptor but will require platelet δ granule secretion (with the release of ADP and TxA2) to obtain full platelet activation. Platelet ATP secretion from δ granules was measured after platelet activation with collagen and ADP that stimulate different pathways to result in the release of ATP from dense granules that was measured using a lumino-aggregometer. Serotonin was measured in plasma after isolation of platelets by centrifugation. It is known that serotonin can active it G protein-coupled receptor but it is a weak agonist that is not able to induce platelet aggregation without stimulation with another agonist. Normal plasma concentrations of serotonin are unable to result in full platelet aggregation. Additional file 2: Figure S2 Platelet aggregation with arachidonic acid. Platelet aggregation in response to 1 mM AA in 159 ASD patients, 103 siblings, 186 parents, and 41 adult controls measured in platelet-rich plasma. Graphs indicating means and 95 % CI value. (PDF 97 kb

Additional file 3: of Newborn genome-wide DNA methylation in association with pregnancy anxiety reveals a potential role for GABBR1

Top 50 DMRcate results. (XLSX 17 kb

PACAP and ceruloplasmin protein levels in nephrin depleted zebrafish.

<p>(A) Western blots for PACAP, ceruloplasmin, and β-actin (loading control) were performed using whole zebrafish lysates at 3 dpf for nephrin depleted (100 μM Nephrin morpholino) compared to control embryos. (B) Signal intensity of Western blot bands was measured using ImageJ software. Graphs represent means ± SD from two repeated experiments. A representative blot is shown. Means were compared using a two-tailed unpaired Student <i>t</i> test, but no significant difference was observed. Mo, nephrin morpholino injected; Co, control morpholino injected.</p

Quantification of thrombocytes in nephrin depleted <i>Tg(cd41</i>:<i>EGFP)</i> transgenic zebrafish.

<p>(A) LEFT: GFP-labeled thrombocytes and thrombocyte precursors are formed in the zebrafish caudal hematopoietic tissue (CHT) region (white arrows) at 3 dpf. Representative pictures of the CHT region of a control and a nephrin depleted (100 μM nephrin morpholino) embryo at 3 dpf are shown. No obvious differences in thrombocyte numbers were observed. RIGHT: Pixel intensity was measured using ImageJ software. The graph represents means ± SD from measurements in three embryos per condition. (B) LEFT: Western blot for GFP and β-actin (loading control) was performed using total zebrafish lysates at 3 dpf. A representative blot is shown. RIGHT: Signal intensity was measured using ImageJ software. The graph represents means ± SD from measurements in two repeated experiments. (C) LEFT: Fluorescence-activated cell sorter (FACS) analysis of control zebrafish lysates for CD41 positive cells was performed at 3 dpf. MIDDLE: FACS analysis of morphant zebrafish lysates for CD41 positive cells was performed at 3 dpf. RIGHT: A diagrammatic representation of the number of GFP-positive cells per 100,000 counted cells. For each zebrafish lysate, 500,000 cells were counted and analyzed. Graphs represent means ± SD from three repeated experiments performed in duplicate. Mo, nephrin morpholino injected; Co, control morpholino injected; SCC, side scatter; GFP, green fluorescent protein.</p

PACAP morpholino suppression and PACAP-38 rescue in nephrin depleted embryos.

<p>(A) Representative larvae of the phenotypes observed in different groups with or without PACAP (<i>adcyap1a and adcyap1b</i>) morpholinos injections (100 μM each). (B) Approximately 100 injected embryos per condition were live-screened at 4 dpf and assigned to a phenotype category. Compared to the control morpholino injection alone, PACAP morpholinos produced a harmful effect when injected together with the control morpholino and a devastating effect with the <i>nphs1</i> morpholino. (C) Representative larvae of the phenotypes observed in different groups with human PACAP-38 injection (5μM). (D) Approximately 100 injected embryos per condition were live-screened at 4 dpf and assigned to a phenotype category. Human PACAP-38 could rescue to a great extent the PACAP morpholino injected embryos; however, they produced no beneficial effect on the <i>nphs1</i> morpholino injected embryos.</p

Quantification of thrombocytes in adriamycin treated zebrafish.

<p>(A) LEFT: GFP-labeled thrombocytes and thrombocyte precursors are formed in the zebrafish caudal hematopoietic tissue (CHT). Representative pictures of the CHT region at 4 dpf of embryos exposed to adriamycin 0, 10 and 30 μM are shown. No obvious difference in thrombocytes was observed between the controls and the adriamycin exposed fish. RIGHT: Pixel intensity was measured using ImageJ software. The graph represents means ± SD from measurements in three embryos per condition. No significant difference was observed between adriamycin exposed and control embryos. (B) LEFT: Western blot for GFP and β-actin (loading control) was performed using total zebrafish lysates at 4 dpf. A representative blot is shown. RIGHT: Signal intensity was measured using ImageJ software. The graph represents means ± SD from measurements in two repeated experiments. No significant difference was observed between adriamycin exposed and control embryos.</p