Journal: Tissue engineering. Part A

Article Title: Developing vasculature and stroma in engineered human myocardium.

doi: 10.1089/ten.TEA.2010.0557

Figure Lengend Snippet: FIG. 3. Endothelial cell net- work formation varies with clonal hMSC lines HS-5 and HS-27a in bi-cell patches. An antibody against rabbit CD31 marked endothelial cells (brown) throughout 8 days of in vitro culture for two-cell patches with HUVECs and either HS-5 or HS-27a hMSCs (A). Endothelial cell networks were more abundant in H: HS-27a patches compared to H:HS-5 patches at all time points. Further, the number of vessel-like structures per mm2

Article Snippet: For identifying rat versus human endothelium in heart sections, sections were blocked with 1.5% normal goat serum in PBS and rat endothelial cell antigen (RECA; mouse anti-RECA-1, 1:10 dilution; Novus) primary antibody was used with sheep anti-mouse HRP IgG secondary antibody (1:100; GE Healthcare) and Tyramide Signal Amplification Kit with Alexa 488 (Invitrogen).

Techniques: In Vitro

Journal: Tissue engineering. Part A

Article Title: Developing vasculature and stroma in engineered human myocardium.

doi: 10.1089/ten.TEA.2010.0557

Figure Lengend Snippet: FIG. 5. In vivo implantation of tri-cell patches on an uninjured rat heart shows human vessel formation is predicted by the presence of endothelial cell networks in vitro. Tri-cell patches with either HS-5 or HS-27a hMSCs were sutured on the epicardial surface of an uninjured athymic rat heart. An antibody to hCD31 (brown) demonstrates few human endothelial cells and no lumen structures in C:H:HS-5 patches (A, B). In contrast, hCD31-positive endothelial cells and lumens are numerous in C:H:HS-27a patches (C, D). Double staining with RECA (red) and hCD31 (green) antibodies demonstrates primarily rat lumens in C:H:HS-5 patches (E) and rat, human, or chimeric lumens in C:H:HS-27a patches (F, G) and in C:H:primary hMSC patches (H). The distribution of lumen diameters was comparable between C:H:HS-27a and C:H:primary hMSC patches, with all lumens being <50 mm (I). The number of hCD31-positive lumens was not different between C:H:HS- 27a and C:H:primary hMSC patches (J). The presence of red blood cells marked by TER-119 (red) in hCD31-positive lumens (green) in C:H:HS-27a patches (K) and C:H:primary hMSC patches (L) demonstrates perfusion from the host. RECA, rat endothelial cell antigen.

Article Snippet: For identifying rat versus human endothelium in heart sections, sections were blocked with 1.5% normal goat serum in PBS and rat endothelial cell antigen (RECA; mouse anti-RECA-1, 1:10 dilution; Novus) primary antibody was used with sheep anti-mouse HRP IgG secondary antibody (1:100; GE Healthcare) and Tyramide Signal Amplification Kit with Alexa 488 (Invitrogen).

Techniques: In Vivo, In Vitro, Double Staining

Journal: European radiology

Article Title: Magnetic resonance imaging for monitoring the effects of thalidomide on experimental human breast cancers

doi: 10.1007/s00330-008-1111-x

Figure Lengend Snippet: Representative immunohistochemically stained human breast cancer sections (MDA-MB-435) showing leakage of macromolecular contrast medium (streptavidin-biotin reaction), vascular richness (lectin and RECA-1 antibody), and VEGF in tumor cells. Thalidomide treatment reduces the extravasation of albumin-(Gd-DTPA)27-(biotin)11 without reducing the abundance of tumor blood vessels. a Tumor section from the saline-control group administered albumin-(Gd-DTPA)27-(biotin)11. The strong red signal indicates extravascular 1-h accumulation of biotin-labeled contrast medium surrounding the yellow-green tumor microvessels. b After 7 days of treatment with thalidomide, the density of red-fluorescent, biotin-labeled contrast agent extravasated over 1 h is strongly reduced compared with a, indicating a reduction in leakage and extravascular accumulation of the macromolecules. Confocal microscopic images of tumor vessels in MDA-MB-435 tumors after treatment with saline (c, d) or thalidomide (e, f) for 7 days show no noticeable change in the area density of perfused blood vessels (green lectin-stained) or total blood vessels (red, RECA-1 stained). No difference is observable (c–f) between saline-control and thalidomide-treated groups with regard to tumor vascularity. (g, h). Representative MDA-MB-435 tumor sections after immunohistochemical staining for human VEGF after a 7-day, three-injection treatment protocol with saline (g) or thalidomide (h). No difference in amount VEGF immunoreactivity was detected in the two groups. Scale bar 115 μm in c–f and 120 μm in a, b

Article Snippet: Endothelial cells, whether lectin-stained or not, were stained with rat monoclonal anti-RECA-1 (1:250, Pharmingen, San Diego, Calif.) to provide a means to assess the entire population of endothelial cells in the section, regardless of their perfusion status.

Techniques: Staining, Labeling, Immunohistochemical staining, Injection

Journal:

Article Title: Molecular Cloning, Nucleotide Sequence, and Expression of Genes Encoding a Polycyclic Aromatic Ring Dioxygenase from Mycobacterium sp. Strain PYR-1

doi: 10.1128/AEM.67.8.3577-3585.2001

Figure Lengend Snippet: Bacterial strains and plasmids

Article Snippet: E. coli JM 109 , Competent cells; F′ proAB lacl q Z Δ M15 , Promega.

Techniques: Plasmid Preparation, In Vivo, Expressing

Journal:

Article Title: Screening of Thermotolerant Gluconobacter Strains for Production of 5-Keto- d -Gluconic Acid and Disruption of Flavin Adenine Dinucleotide-Containing d -Gluconate Dehydrogenase ▿

doi: 10.1128/AEM.00640-09

Figure Lengend Snippet: Bacterial strains, plasmids, and primers used in this study

Article Snippet: The oligonucleotides used in this study were purchased from FASMAC Co. Ltd. (Kanagawa, Japan) and are listed in Table . table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Strain, plasmid, or primer Relevant trait(s) or sequence Source or reference E. coli DH5α r − m + recA1 lacZYA φ80d lac Δ( lacZ )M15 Bethesda Research Laboratories, Inc. Gluconobacter strains THE42 Parental strain 12 THE42 gndG ::Km Km r , strain THE42 with gndG disrupted This study THF55 Parental strain 12 THF55 gndG ::Km Km r , strain THF55 with gndG disrupted This study THG42 Parental strain 12 THG42 gndG ::Km Km r , strain THG42 with gndG disrupted This study Plasmids pGEM-T Easy Cloning vector Promega pTKm Contains nonpolar Km r cassette 23 pGEM-CA-F 1.9-kb PCR product inserted into pGEM-T Easy This study pGEM-CA-F::Km Km r , used for gndG disruption, Km r cassette inserted at SmaI site This study Primers gndL-F 5′-CCGTCAACTCGCAGCCTTTCTGC-3′ This study gndL-R 5′-GCGGGCAGGATATTCACGTTGGG-3′ This study F-igL-1 5′-TAGGTGACGCCGTAATCCTG-3′ This study R-igL-2 5′-TCGGATACCATCCCTATTCCC-3′ This study F-gnDH-3 5′-TCATGGACTGGGTCAATCAG-3′ This study R-gnDH-2 5′-TGCAAGCAAAGGCCAGGATT-3′ This study Open in a separate window Bacterial strains, plasmids, and primers used in this study Bacterial strains and growth conditions.

Techniques: Plasmid Preparation, Sequencing, Clone Assay