Parasite cell division depends on a fiber that once anchored the basal body of the flagellum in the algal ancestor. Here, you see the fiber (green), centrosomes (red), parasite daughter cells (blue), and nucleus (grey). The micrograph on the right depicts two <i>Toxoplasma gondii</i> parasites in division.

<p>Parasite cell division depends on a fiber that once anchored the basal body of the flagellum in the algal ancestor. Here, you see the fiber (green), centrosomes (red), parasite daughter cells (blue), and nucleus (grey). The micrograph on the right depicts two <i>Toxoplasma gondii</i> parasites in division.</p

Supplemental Material for Zhang et al., 2018

<p><a><b>Figure S1 RNAi knockdown of Class 3 and 4 genes led to defects in oocyte development</b></a>. Ovaries dissected from 3-5 days old control or knockdown females were stained for DNA (red) using propidium iodide. A summary of the ovarian phenotypes can be found in Table S3. Images are arranged according to observed stage of oogenesis defect, from early stage to late stage. Scale bar: 50 µm.</p> <p> </p> <p><a><b>Figure S2. Maternal knockdown of 12 genes led to reduction and elimination of egg hatchability that recapitulated previously reported maternal effect phenotypes.</b></a> 2-4 hr old embryos from knockdown females, stained for tubulin (green) and DNA (red). For each set of images, the image on the right shows a close-up view of the spindle (marked by the arrow on the left). Images are organized in alphabetical order. The AttP2 panel, the control for comparison, is the same as in Fig. 2. Scale bar: oocyte image 50 µm, enlarged image 10 µm.</p> <p> </p> <p><a><b>Figure S3.</b></a> <b>Fertilization was not significantly impacted in the early arrest knockdown embryos of the 27 genes</b> Sperm tail (green; arrow) was detected in the stage 1-2 arrested embryos produced by RNAi females for 27 genes. The images are ordered in alphabetical order, by gene name. Scale bar: 50 µm.</p> <p> </p> <p><a><b>Figure S4. Translation of <i>smg</i> in unfertilized eggs produced by control and RNAi females of 27 early maternal effect candidates</b></a>. Among the 27 genes examined, Smg protein was detected in the activated eggs produced by RNAi females of 26 genes. Translation of Smg protein was not detected in the activated eggs produced by <i>plu</i> knockdown females.<br><br><b>Table S1</b> <b>- S7</b><br></p

Additional file 1: of Physical activity and risk of testicular cancer: a systematic review

Table S1. Papers included in meta-analysis of association between physical activity and testicular cancer risk, with study meta-data. Table S2. Extracted data relating to high vs. low physical activity. Table S3. Extracted data relating to high vs. low recreational physical activity at adolescence/early adulthood. Table S4. PICOS (Patient/Participant, Intervention, Comparator, Outcome, Study design) criteria for inclusion of studies. Table S5. List of excluded papers with reason for exclusion. Table S6. Assessment of study quality against Newcastle-Ottawa criteria for case-control studies. Table S7. Assessment of study quality against Newcastle-Ottawa criteria for cohort studies. (DOCX 134 kb

Integrative Bioinformatics Links HNF1B with Clear Cell Carcinoma and Tumor-Associated Thrombosis

<div><p>Clear cell carcinoma (CCC) is a histologically distinct carcinoma subtype that arises in several organ systems and is marked by cytoplasmic clearing, attributed to abundant intracellular glycogen. Previously, transcription factor hepatocyte nuclear factor 1-beta (HNF1B) was identified as a biomarker of ovarian CCC. Here, we set out to explore more broadly the relation between HNF1B and carcinomas with clear cell histology. HNF1B expression, evaluated by immunohistochemistry, was significantly associated with clear cell histology across diverse gynecologic and renal carcinomas (<i>P</i><0.001), as was hypomethylation of the <i>HNF1B</i> promoter (<i>P</i><0.001). From microarray analysis, an empirically-derived HNF1B signature was significantly enriched for computationally-predicted targets (with HNF1 binding sites) (<i>P</i><0.03), as well as genes associated with glycogen metabolism, including glucose-6-phophatase, and strikingly the blood clotting cascade, including fibrinogen, prothrombin and factor XIII. Enrichment of the clotting cascade was also evident in microarray data from ovarian CCC <i>versus</i> other histotypes (<i>P</i><0.01), and HNF1B-associated prothrombin expression was verified by immunohistochemistry (<i>P</i> = 0.015). Finally, among gynecologic carcinomas with cytoplasmic clearing, HNF1B immunostaining was linked to a 3.0-fold increased risk of clinically-significant venous thrombosis (<i>P</i> = 0.043), and with a 2.3-fold increased risk (<i>P</i> = 0.011) in a combined gynecologic and renal carcinoma cohort. Our results define HNF1B as a broad marker of clear cell phenotype, and support a mechanistic link to glycogen accumulation and thrombosis, possibly reflecting (for gynecologic CCC) derivation from secretory endometrium. Our findings also implicate a novel mechanism of tumor-associated thrombosis (a major cause of cancer mortality), based on the direct production of clotting factors by cancer cells.</p></div

Nuclear HNF1B expression is associated with cytoplasmic clearing across multiple tumor types.

<p>Representative H&E (10×objective, and magnified inset) and positive HNF1B immunostains are shown for (<b>A</b>) ovarian CCC, (<b>B</b>) renal CCC, and (<b>C</b>) mixed endometrioid/clear cell ovarian carcinoma. In the ovarian carcinoma with mixed histology (C), the upper row of panels depicts a region of the tumor with clear cell histology (HNF1B-positive); the middle row of panels (insets) depicts a different region of the same tumor with endometrioid histology and clear cell features (note papillary pattern, highlighted by green box) (also HNF1B-positive); and the bottom row (insets) depicts yet another region of the same tumor with endometrioid histology but without clear cell features (and with correspondingly weaker HNF1B-immunostaining). Note that this patient experienced a tumor-associated thromboembolic event (see main text). (<b>D</b>) Graphical display of proportion of HNF1B positive cases (shaded red) among different carcinoma types with or without cytoplasmic clearing. Gynecologic carcinomas include those from the endometrium, cervix and ovary. Other neoplasms represent diverse anatomic sites (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0074562#pone.0074562.s001" target="_blank">Table S1</a>). <i>P</i>-values (two-sided Fisher’s exact test) for pairwise comparisons are indicated.</p

Tumor HNF1B expression is associated with venous thrombosis.

1<p>Two-sided Fisher’s exact test.</p

HNF1B-positive gynecologic malignancies are associated with cytoplasmic prothrombin expression, and increased clotting factor transcript levels.

<p>(<b>A</b>) Representative prothrombin immunostains (10×objective, and magnified inset) are shown for HNF1B-negative, prothrombin-negative mixed-histology endometrial carcinoma (<i>left</i>); and HNF1B-positive, prothrombin-positive (moderate cytoplasmic staining) mixed endometrioid/clear cell ovarian carcinoma (<i>right</i>; same case as depicted in Fig. 1C). (<b>B</b>) Q-RT-PCR analysis demonstrates increased clotting factor transcript levels in ovarian CCC (n = 6) compared to ovarian serous carcinoma (n = 7). Transcript levels are normalized to GAPDH, then set relative to a single serous carcinoma “reference” sample, and reported as log2 values. For graphs shown, undetected transcript levels (green diamonds) are arbitrarily set to smallest detectable levels. Red bars indicate average levels; <i>P</i>-values (non-parametric Mann-Whitney U-test) are shown.</p

HNF1B transcriptional targets and clotting cascade are enriched in ovarian CCC.

<p>(<b>A</b>) GSEA analysis shows enrichment of putative HNF1 targets (“leading edge” from Fig. 3) among genes selectively expressed in ovarian carcinoma with clear cell <i>vs</i>. other histotype <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0074562#pone.0074562-Hendrix1" target="_blank">[25]</a>. (<b>B</b>) The leading edge enriched genes determined here (n = 25) are significantly over-represented among select functional gene sets (nominal <i>P</i> values<0.01); the top 10 gene sets are shown. Columns on the left depict relative log₂ gene expression levels (red/blue scale indicated) among normal ovary and ovarian carcinomas of different histotype. Columns on the right indicate membership (gray fill) among the top 10 most significant gene sets, which include liver-specific genes, and the clotting cascade (represented by five different gene sets). (<b>C</b>) Clotting factor genes (<i>FGA</i>, <i>FGB</i>, <i>F2</i>, and <i>F13B</i>) are more highly expressed in laser-microdissected ovarian CCC <i>vs</i>. normal ovarian surface epithelium (dataset from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0074562#pone.0074562-Stany1" target="_blank">[26]</a>). Heatmap depicts all microarray probes for the respective genes; corresponding <i>P</i>-values (two-sided Student’s t-test) are indicated.</p