Journal: Journal of Fungi

Article Title: Chromogenic, Biochemical and Proteomic Identification of Yeast and Yeast-like Microorganisms Isolated from Clinical Samples from Animals of Costa Rica

doi: 10.3390/jof10030218

Figure Lengend Snippet: Results of the identification of the isolates by Brilliance ® Candida , Vitek ® 2 Compact and MALDI-TOF Biotyper ® MSP.

Article Snippet: Proteomic identification by mass spectrophotometry with MALDI-TOF was performed using the MALDI Biotyper ® MSP (Bruker Daltonics GmBH & Co. KG, Bremen, Germany) from the Mycology Reference Laboratory of the Costa Rican Institute for Research and Teaching in Nutrition and Health (INCIENSA) to corroborate the biochemical identification.

Techniques: Cream

Journal: Frontiers in Endocrinology

Article Title: When RON MET TAM in Mesothelioma: All Druggable for One, and One Drug for All?

doi: 10.3389/fendo.2019.00089

Figure Lengend Snippet: MST1R (RON) is activated in MPM patient samples and cell lines. (A) A heat map summarizing the basal phosphorylation levels of the MET (HGFR), MST1R (RON), and the TAM RTKs (TYRO3, AXL, and MERTK) in MPM tumors ( n = 7) and cell lines ( n = 4; Ju77, NCI-H28, NCI-H2052, ONE58). Signals with an intensity value greater than the 99% confidence interval of the mean of the 10 negative controls were scored as positive. Yellow indicates high activity and blue indicates low or undetectable kinase activity. (B) flMST1R and sfMST1R, MET, MST1, AXL, TYRO3, MERTK, and GAS6 were detected at the mRNA level (standard end point PCR), in a panel of MPM cell lines, which included two normal mesothelial cell lines (LP9 and Met5A) ( n = 17). 18S rRNA was used as a loading control.

Article Snippet: Recombinant human MST1 was obtained from R & D Systems (MN, USA) and re-suspended in sterile PBS containing 0.1% BSA w/v.

Techniques: Phospho-proteomics, Activity Assay, Control

Journal: Frontiers in Endocrinology

Article Title: When RON MET TAM in Mesothelioma: All Druggable for One, and One Drug for All?

doi: 10.3389/fendo.2019.00089

Figure Lengend Snippet: Levels of MST1 are significantly elevated in MPM tissue vs. benign. (A) MST1 is overexpressed in primary MPM vs. benign pleura. Analysis of the Leicester samples showed significant ( p = 0.0022) overexpression in tumors ( n = 17) vs. benign pleura ( n = 5) Statistical significance was obtained using a two tailed Student's t -test with Welch's correction (** p < 0.01). This data was confirmed using Oncomine analysis of the Gordon MPM dataset ( n = 49) , where MST1 levels were elevated in MPM tumors compared to pleural tissue ( p = 0.018). For differential gene expression analysis Oncomine uses two sided Student's t -test . (B) MST1 is expressed at protein level in MPM cell lines ( n = 12). Western blot demonstrating expression of MST1 at the protein level in MPM cell lines. (C) Serum MST1 levels were unaltered in patients with primary mesothelioma vs. patients with prior exposure to asbestos ( n = 20 per group). (D) GAS6 mRNA levels were unchanged in the MPM patient cohort compared with benign pleura. β-actin or 18S rRNA is included as a loading control in appropriate images.

Article Snippet: Recombinant human MST1 was obtained from R & D Systems (MN, USA) and re-suspended in sterile PBS containing 0.1% BSA w/v.

Techniques: Over Expression, Two Tailed Test, Gene Expression, Western Blot, Expressing, Control

Journal: Frontiers in Endocrinology

Article Title: When RON MET TAM in Mesothelioma: All Druggable for One, and One Drug for All?

doi: 10.3389/fendo.2019.00089

Figure Lengend Snippet: In a cohort of MPM patient samples, elevated levels of MST1R (RON) and MST1 are associated with increased survival. (A) Immunohistochemical staining of an MPM TMA for MST1R (Santa Cruz Biotechnology, sc-322, RRID: AB_677390 ), at (40x) magnification for both focal positive and strongly positive cores. Following scoring by two pathologists, and dichotomized closest to the median into low and high using >2, 2, = 10%, Kaplan-Meier Analyses (with OS from time of diagnosis) was not significant (14.5 vs. 6.6 months p = 0.052).

Article Snippet: Recombinant human MST1 was obtained from R & D Systems (MN, USA) and re-suspended in sterile PBS containing 0.1% BSA w/v.

Techniques: Immunohistochemical staining, Staining, Expressing, Negative Control, Biomarker Discovery

Journal: Frontiers in Endocrinology

Article Title: When RON MET TAM in Mesothelioma: All Druggable for One, and One Drug for All?

doi: 10.3389/fendo.2019.00089

Figure Lengend Snippet: Treatment with LCRF-0004 and BMS-77607 reduces the proliferative capacity of malignant mesothelioma cells. (A) Effects of LCRF-0004 (200 nM) and MST1 (250 ng/mL) (alone or in combination) on cellular proliferation as measured using a BrdU proliferation assay. LCRF-0004 significantly affects the proliferative capacity of NCI-H226 cells at 24 or 48 h, while MST1 is unable to rescue the cells from the effect of RON inhibition. (B) Cellular viability is also decreased in response to LCRF-0004 treatment (HCS assay) using the NCI-H226 cells. (C) Treatment with BMS-77607 resulted in significantly reduced cellular proliferation in both NCI-H226 and REN (MPM cell lines), whilst having no effect on LP9 (normal mesothelial cells). Significance was calculated based on a one-way ANOVA with a post-hoc Tukey's Multiple Comparison test (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: Recombinant human MST1 was obtained from R & D Systems (MN, USA) and re-suspended in sterile PBS containing 0.1% BSA w/v.

Techniques: Proliferation Assay, Inhibition, Comparison

Journal: Frontiers in Endocrinology

Article Title: When RON MET TAM in Mesothelioma: All Druggable for One, and One Drug for All?

doi: 10.3389/fendo.2019.00089

Figure Lengend Snippet: LCRF-0004 promotes apoptosis and alters cell cycle progression in MPM cells (A) Inhibition of MST1R (RON) by LCRF-0004 (200 nM) promotes cellular apoptosis in the NCI-H226 cell lines as measured by FACS (48 h post treatment). A representative FACS plots are show with the results graphed underneath (MST1–250 ng/mL). Significance was calculated based on a one-way ANOVA with a post-hoc Tukey's Multiple Comparison test. (** p < 0.01; *** p < 0.001). (B) FACS analysis determined that LCRF-0004 (200 nM) treatments result in a significant accumulation of cells in the G2/M phase. Significance based on a one-way ANOVA with a Tukey's Multiple Comparisons test (** p < 0.01).

Article Snippet: Recombinant human MST1 was obtained from R & D Systems (MN, USA) and re-suspended in sterile PBS containing 0.1% BSA w/v.

Techniques: Inhibition, Comparison

Journal: Frontiers in Endocrinology

Article Title: When RON MET TAM in Mesothelioma: All Druggable for One, and One Drug for All?

doi: 10.3389/fendo.2019.00089

Figure Lengend Snippet: Primers and associated annealing temperatures.

Article Snippet: Recombinant human MST1 was obtained from R & D Systems (MN, USA) and re-suspended in sterile PBS containing 0.1% BSA w/v.

Techniques: Sequencing, Mutagenesis

Journal: International Journal of Molecular Sciences

Article Title: The Variation Analysis of DNA Methylation in Wheat Carrying Gametocidal Chromosome 3C from Aegilops triuncialis

doi: 10.3390/ijms18081738

Figure Lengend Snippet: Representative variation of DNA methylation pattern. H ( Eco R I + Hpa II digest) and M ( Eco R I + Msp I digest) refer to digestion with Eco R I + Hpa II and Eco R I + Msp I, respectively. “→” red arrows represent parts of differential methylated bands between common wheat and wheat carrying Gc chromosome(s). M stands for Marker DL2000. MA (1, 1), presence in both H ( Eco R I + Hpa II digest) and M ( Eco R I + Msp I digest) lanes; MB (1, 0), presence in H and absence in M lane; MC (0, 1), absence in H but presence in M lane; MD (0, 0) absence in both H and M lanes. CS: T. aestivum cv. Chinese Spring. CS–3C: monosomic addition line of Chinese Spring (CS) that carries a gametocidal chromosome 3C originated from Aegilops triuncialis . CS–3C3C: disomic addition line of Chinese Spring (CS) that carries two gametocidal chromosome 3C originated from Aegilops triuncialis .

Article Snippet: The Genomic DNA sample was digested by Eco R I/ Hpa II and Eco R I/ Msp I independently and simultaneously at 37 °C for 6 h and then ligated with Eco R I and Hpa II/ Msp I adapters ( ) at 8 °C for 4 h. The digestion and ligation reaction system was in a total volume of 20 μL containing 3 μL genomic DNA, 1 μL Eco R I (R0101L, NEB), 1 μL Hpa II (R0171L, NEB) or Msp I (R0106L, NEB), 1 μL Eco R I adapter, 1 μL Hpa II/ Msp I adapters, 1 μL T4 DNA ligase (R0202L, NEB), 2 μL 10× T4 buffer (R0202L, NEB) and double-distilled water to 20 μL.

Techniques: DNA Methylation Assay, Methylation, Marker