Ultrasonographic assessment of costochondral cartilage for microtia reconstruction

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/149363/1/lary27390_am.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/149363/2/lary27390.pd

Melanoma cells break down LPA to establish local gradients that drive chemotactic dispersal.

The high mortality of melanoma is caused by rapid spread of cancer cells, which occurs unusually early in tumour evolution. Unlike most solid tumours, thickness rather than cytological markers or differentiation is the best guide to metastatic potential. Multiple stimuli that drive melanoma cell migration have been described, but it is not clear which are responsible for invasion, nor if chemotactic gradients exist in real tumours. In a chamber-based assay for melanoma dispersal, we find that cells migrate efficiently away from one another, even in initially homogeneous medium. This dispersal is driven by positive chemotaxis rather than chemorepulsion or contact inhibition. The principal chemoattractant, unexpectedly active across all tumour stages, is the lipid agonist lysophosphatidic acid (LPA) acting through the LPA receptor LPAR1. LPA induces chemotaxis of remarkable accuracy, and is both necessary and sufficient for chemotaxis and invasion in 2-D and 3-D assays. Growth factors, often described as tumour attractants, cause negligible chemotaxis themselves, but potentiate chemotaxis to LPA. Cells rapidly break down LPA present at substantial levels in culture medium and normal skin to generate outward-facing gradients. We measure LPA gradients across the margins of melanomas in vivo, confirming the physiological importance of our results. We conclude that LPA chemotaxis provides a strong drive for melanoma cells to invade outwards. Cells create their own gradients by acting as a sink, breaking down locally present LPA, and thus forming a gradient that is low in the tumour and high in the surrounding areas. The key step is not acquisition of sensitivity to the chemoattractant, but rather the tumour growing to break down enough LPA to form a gradient. Thus the stimulus that drives cell dispersal is not the presence of LPA itself, but the self-generated, outward-directed gradient

Quality control and quantification in IG/TR next-generation sequencing marker identification: protocols and bioinformatic functionalities by EuroClonality-NGS

Assessment of clonality, marker identification and measurement of minimal residual disease (MRD) of immunoglobulin (IG) and T cell receptor (TR) gene rearrangements in lymphoid neoplasms using next-generation sequencing (NGS) is currently under intensive development for use in clinical diagnostics. So far, however, there is a lack of suitable quality control (QC) options with regard to standardisation and quality metrics to ensure robust clinical application of such approaches. The EuroClonality-NGS Working Group has therefore established two types of QCs to accompany the NGS-based IG/TR assays. First, a central polytarget QC (cPT-QC) is used to monitor the primer performance of each of the EuroClonality multiplex NGS assays; second, a standardised human cell line-based DNA control is spiked into each patient DNA sample to work as a central in-tube QC and calibrator for MRD quantification (cIT-QC). Having integrated those two reference standards in the ARResT/Interrogate bioinformatic platform, EuroClonality-NGS provides a complete protocol for standardised IG/TR gene rearrangement analysis by NGS with high reproducibility, accuracy and precision for valid marker identification and quantification in diagnostics of lymphoid malignancies.This work was supported by Ministry of Health of the Czech Republic, grant no. 16-34272A; computational resources were provided by the CESNET LM2015042 and the CERIT Scientific Cloud LM2015085, provided under the programme “Projects of Large Research, Development, and Innovations Infrastructures”. Analyses in Prague (JT, EF and MS) were supported by Ministry of Health, Czech Republic, grant no. 00064203, and by PRIMUS/17/MED/11. Analyses in the Monza (Centro Ricerca Tettamanti, SS, AG and GC) laboratory were supported by the Italian Association for Cancer Research (AIRC) and Comitato Maria Letizia Verga

Left Hemisphere Specialization for Oro-Facial Movements of Learned Vocal Signals by Captive Chimpanzees

The left hemisphere of the human brain is dominant in the production of speech and signed language. Whether similar lateralization of function for communicative signal production is present in other primates remains a topic of considerable debate. In the current study, we examined whether oro-facial movements associated with the production of learned attention-getting sounds are differentially lateralized compared to facial expressions associated with the production of species-typical emotional vocalizations in chimpanzees.Still images captured from digital video were used to quantify oro-facial asymmetries in the production of two attention-getting sounds and two species-typical vocalizations in a sample of captive chimpanzees. Comparisons of mouth asymmetries during production of these sounds revealed significant rightward biased asymmetries for the attention-getting sounds and significant leftward biased asymmetries for the species-typical sounds.These results suggest that the motor control of oro-facial movements associated with the production of learned sounds is lateralized to the left hemisphere in chimpanzees. Furthermore, the findings suggest that the antecedents for lateralization of human speech may have been present in the common ancestor of chimpanzees and humans approximately 5 mya and are not unique to the human lineage

mRNA Coronavirus Disease 2019 Vaccine-Associated Myopericarditis in Adolescents: A Survey Study

In this survey study of institutions across the US, marked variability in evaluation, treatment, and follow-up of adolescents 12 through 18 years of age with mRNA coronavirus disease 2019 (COVID-19) vaccine-associated myopericarditis was noted. Only one adolescent with life-threatening complications was reported, with no deaths at any of the participating institutions

Growth factors potentiate LPA chemotaxis.

<p>(A) Growth factors enhance cells' response to LPA gradients. Figure shows plots of the WM239A paths chemotaxing in gradients of LPA, LPA+EGF+PDGF, and conflicting gradients of LPA versus EGF+PDGF. (B) Chemotactic indices of cells in (A) and other conditions. Growth factor gradients if anything increase the efficiency of LPA chemotaxis, even when applied in a gradient in the opposite direction. Bars show SEM.</p

LPA gradients across melanomas <i>in vivo</i>.

<p>(A) TYR::CreER^T2BRAF^V600E/+PTEN^lox/+ mice, a genetically appropriate melanoma model, were treated with tamoxifen as described in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001966#pbio.1001966-Garcia1" target="_blank">[56]</a>, grown until melanomas spontaneously developed. Dashed box shows the region used for the samples shown in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001966#pbio-1001966-g001" target="_blank">Figure 1C</a>. (B) Haematoxylin and eosin-stained biopsies of murine melanomas demonstrating the dispersal of cells from a representative tumour from <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001966#pbio-1001966-g005" target="_blank">Figure 5A</a>, with cells spreading directly away from the tumour. Upper image 2.5× magnification; lower image 20× magnification from dashed box above, showing melanoma cells invading toward the muscle layer (D, dermis; M, muscle layer). (C) Biopsies from mouse melanomas. Several sites in a linear distribution were biopsied using a 6 mm punch biopsy tool within 5 minutes of the mouse being sacrificed and immediately frozen in liquid nitrogen. The positions of biopsies used for LPA measurement are indicated (too few distant samples were obtained for a significant measurement). Bar shows 5 mm. (D) LPA concentration gradients across the margin of a melanoma. Four melanomas were sampled at three sites in a line as shown in (A) (A, tumour body; B, tumour edge; C, skin surrounding tumour). Total LPA per mg tissue was quantified by mass spectrometry after weighing the tissue specimens and extracting the LPA. Outward-directed gradients of LPA were found across the margin of all the melanomas tested. Bars show SEM. (E) Analysis of LPA subspecies. 18∶2-LPA, 20∶4-LPA and 22∶6-LPA show a clearer gradient than 16∶0-LPA, which is though to be less active as a signalling molecule.</p

Dispersal is due to a chemoattractant present in serum.

<p>All panels show data from melanoma cells migrating in chemotaxis chambers as described <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001966#pbio.1001966-MuinonenMartin1" target="_blank">[15]</a>. (<b>A–B</b>) Cells migrate from conditioned medium towards fresh medium. WM1158 cells were randomly attached to a coverslip and assembled in a chamber in 48 hour WM1158 cell conditioned medium. The medium in one chamber was replaced with fresh medium, while the other was left alone. Tracks of individual cells are shown as coloured lines (A). Cells move towards the fresh medium, as shown by the spider plot (B) showing all cell tracks. (<b>C–D</b>) Example images showing WM239A metastatic melanoma cells after 21 hours in serum-free medium (C) and a 0%–10% FBS gradient (D). Coloured paths show centroid tracks from time 0. (<b>E</b>) Quantitative analysis of chemotactic responses. “Spider” plots (large panels), rose plots, mean chemotactic index, and Rayleigh test for directionality are shown for cells in serum-free medium and a 0%–10% FBS gradient (<i>n</i>>100 cells in three independent experiments for both conditions). Spider plots show strong chemotaxis in FBS gradients; in serum-free medium only random movement is seen. Rose plots show overall movement from 6–12 hours; the proportion of total cells in each sector is shown on a log scale, with red lines representing the 95% confidence interval. The majority of cells in the FBS gradient move in the direction of the chemoattractant. Rayleigh tests statistically confirmed this highly significant unimodal directionality. Graphs of chemotactic index were generated from the same data.</p

Chemotaxis of cells from different melanoma stages.

<p>(A) Chemotaxis of a panel of six cell lines from different melanoma stages (RGP, green; VGP, purple; metastatic, red) up a 0%–10% FBS gradient was measured as above (<i>n</i>≥45 cells per cell line). (B) Chemotactic index of cells from different stages. Data from (A) were collated by melanoma stage. Chemotaxis improves as the stage of melanoma progresses, although even the earliest RGP cells show clear chemotaxis. (C) Speeds of cells from different stages. Data from (A) were collated by melanoma stage. Metastatic lines are conspicuously faster (<i>p</i>-values from unpaired <i>t</i>-tests), although again the speed of RGP and VGP cells is still relatively high for non-haematopoietic cells.</p

Density-dependent dispersal of melanoma cells.

<p>(A) Schematic showing the stages of melanoma spread. (B) WM239A metastatic melanoma cells dispersing in uniform medium. 2×10⁴ cells were introduced into one reservoir of an Insall chamber containing complete medium with 10% FBS throughout, and observed by time-lapse phase contrast microscopy. See <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001966#pbio.1001966.s004" target="_blank">Movie S1</a>. The left side of each image shows the reservoir containing cells, while the right side is the viewing bridge of the chamber. (C–D) Migration is density-dependent. WM1158 metastatic melanoma cells were seeded at different densities in full medium with 10% FBS, and observed as before. At 2×10⁴ cells/well and above, peak migration distances increase sharply, as confirmed by the distance at 17 hours (D; graph shows mean ± SEM). (E) Migration is not driven by production of a repellent. 2×10⁴ WM1158 cells were introduced into a chamber in minimal medium without serum and observed at 17 hours as before. Cells survive and adhere, but do not disperse. (F) Migration is not driven by production of a serum-derived repellent. 2×10⁴ WM1158 cells were introduced into a chamber in minimal medium without serum and observed at 17 hours as before. Cells disperse less efficiently in conditioned medium than in fresh medium. (G) Migration mediated by chemotaxis up a serum gradient is similar to density-induced migration. Left panel: 2×10⁴ WM1158 cells were introduced into a chamber in the presence of a gradient from 0% FBS around the cells to 10% in the opposite reservoir <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001966#pbio.1001966-MuinonenMartin1" target="_blank">[15]</a>. The cells rapidly migrate towards the well containing serum. Right panel: similar assay with 10% serum in both reservoirs. Panels taken from <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001966#pbio.1001966.s006" target="_blank">Movies S3</a> and <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.1001966#pbio.1001966.s004" target="_blank">S1</a>, respectively.</p