Journal: The Journal of Cell Biology

Article Title: Identification of a putative pathway for the muscle homing of stem cells in a muscular dystrophy model

doi: 10.1083/jcb.200210006

Figure Lengend Snippet: FACscan ® immunophenotyping of the muscle-derived cells isolated using MACS ® columns. The MDSCs obtained by six preplatings were tested for CD34 and Sca-1 antigen expression. Three distinct populations of MDSCs were obtained: Sca-1 + /CD34 − (a, first screen), Sca-1 + /CD34 + (a, second screen), and Sca-1 − /CD34 + (a, third screen). In b, the donor cells were detected by PCR for the Y chromosome in the hind limb muscles (lane 1), blood (lane 2), and bone marrow (lane 3), but not in samples from organs such as the liver (lane 4), spleen (lane 5), lung (lane 6), and brain (lane 7). 3 wk after i.m. transplantation of H2d unselected MDSCs in mdx mice (H2b), engrafted tissues were analyzed by FACS ® for the presence of positive H2d (donor) cells (c). The positive H2d cells from the blood tissue (boxed region indicated in the second quadrant of the blood row) of the injected animals represent ∼0.1% of the total bloodstream. A similar percentage (∼1%) of donor cells was also detected in the bone marrow tissues after 3 wk (c, bone marrow row). We observed <5% donor cells in the controlateral hind limb muscles. The H2d expression in the bloodstream correlates with the expression of Flk-1 and stem cell marker Sca-1, whereas in the bone marrow, donor cells coexpressed the CD90 (12%) and Sca-1 (85%) antigens (c). The muscle tissue also contained some donor cells positive for Flk-1, CD34, and Sca-1 markers. A few donor cells (H2d positive) were also positive for CD34 (45%), Sca-1 (20%), or Flk1 (3%), as indicated by double immunostaining.

Article Snippet: After the isolation of Sca-1 + cells, multisort microbeads were removed using the MACS ® multisort release reagent, and the Sca-1 + cells were incubated with a biotinated, conjugated CD34 antibody (1/50) and resuspended with an antibiotin conjugated with paramagnetic microbeads (Miltenyi Biotec).

Techniques: Derivative Assay, Isolation, Expressing, Muscles, Transplantation Assay, Injection, Marker, Double Immunostaining

Journal: The Journal of Cell Biology

Article Title: Identification of a putative pathway for the muscle homing of stem cells in a muscular dystrophy model

doi: 10.1083/jcb.200210006

Figure Lengend Snippet: We tested the adhesion of the Sca-1 + /CD34 − /L-selectin + positive cells (HMDSCs) to muscle blood vessels by intravital microscopy. Boluses of 5 × 10 5 fluorescently (BCECF) labeled cells were injected into the quadriceps muscle, and their adhesion to the pectoral muscle vessels was recorded. To improve contrast between the intra- and extravascular compartments, the animals were injected intravenously with a low dose of FITC–dextran. The time course study, i.e., before (a), during (b), and after (c and d) i.m. injections, indicates a significant migration of the HMDSCs within the injected muscle. β-Gal staining of ROSA26-derived HMDSCs showed the presence of injected cells along the injection site (e), near myofibers, bleeding vessels (f), and arterioles (g). Moreover, after i.m. injections in quadriceps, the HMDSCs were able to migrate to the blood stream and then adhered to the endothelial lining of several distant muscles, such as pectoralis (h–m). Arrows in i, k, and m indicate migrated BCECF-labeled cells (bright dots) in the perivascular space of a muscle venule 1 h after injection.

Article Snippet: After the isolation of Sca-1 + cells, multisort microbeads were removed using the MACS ® multisort release reagent, and the Sca-1 + cells were incubated with a biotinated, conjugated CD34 antibody (1/50) and resuspended with an antibiotin conjugated with paramagnetic microbeads (Miltenyi Biotec).

Techniques: Intravital Microscopy, Labeling, Injection, Migration, Staining, Derivative Assay, Muscles

Journal: The Journal of Cell Biology

Article Title: Identification of a putative pathway for the muscle homing of stem cells in a muscular dystrophy model

doi: 10.1083/jcb.200210006

Figure Lengend Snippet: Myogenic differentiation of Sca-1 + /CD34 − MDSCs in cultures. Characterization of Sca-1 + /CD34 − MDSCs from Rosa26 and Des-LacZ transgenic mice for the L-selectin expression (a). L-selectin expression by Sca-1 + /CD34 − MDSCs (a, first screen) was decreased when these cells differentiated in myoblasts (a, second screen) and lost when they formed myotubes (a, third screen). For cloning, single cells within the Sca-1 + /CD34 − MDSCs (b, inset) were plated in proliferative conditions described in the Materials and methods section. Sca-1 + /CD34 − /L-selectin + cells (HMDSCs) obtained from clone G13 differentiated well into myotubes (b and c) expressing the late myogenic marker MyHC (d). Few round single cells near myotubes were positive for the expression of m-cadherin (e) and desmin (f) markers. Panel g corresponds to the merging of e and f. Metalloproteinase activity of the HMDSCs (h, lane 2) was also tested and compared with that of myoblasts (h, lane 1). (i) Immunoblotting analysis of slow MyHC and m-cadherin by HMDSCs obtained from clone G13. The first lane of all immunoblottings corresponds to a homogenate of clone G13. 3T3 fibroblast and G8 myoblast cell lines were used as controls. Utrophin immunoblotting indicated that the same total protein concentrations were present in all specimens.

Article Snippet: After the isolation of Sca-1 + cells, multisort microbeads were removed using the MACS ® multisort release reagent, and the Sca-1 + cells were incubated with a biotinated, conjugated CD34 antibody (1/50) and resuspended with an antibiotin conjugated with paramagnetic microbeads (Miltenyi Biotec).

Techniques: Transgenic Assay, Expressing, Cloning, Marker, Activity Assay, Western Blot

Journal: Urologia

Article Title: Introducing in clinical practice a new laser suction handpiece for percutaneous nephrolithotomy

doi: 10.1177/03915603211031874

Figure Lengend Snippet: The Lumenis laser suction handpiece composed of a 11.3 Fr external sheath and a handle with two buttons activated by the surgeon’s thumb.

Article Snippet: Karl Storz later released the LASER Suction Tube ® , which was a simple reusable metallic tube with a side connection to negative pressure, and Lumenis ® presented the LSH as a single-use 11.3 Fr device.

Techniques:

Journal: Urologia

Article Title: Introducing in clinical practice a new laser suction handpiece for percutaneous nephrolithotomy

doi: 10.1177/03915603211031874

Figure Lengend Snippet: Outcome of conventional lithotripsy compared to the use of the laser suction handpiece.

Article Snippet: Karl Storz later released the LASER Suction Tube ® , which was a simple reusable metallic tube with a side connection to negative pressure, and Lumenis ® presented the LSH as a single-use 11.3 Fr device.

Techniques:

Journal: Journal of Pharmaceutical Policy and Practice

Article Title: Drug utilization patterns among elderly hospitalized patients on poly-pharmacy in Punjab, Pakistan

doi: 10.1186/s40545-017-0112-z

Figure Lengend Snippet: Drug utilization pattern in study participants (ATC level 1–4)

Article Snippet: , , , R03: Drugs for obstructive airway diseases , 113 , 18.8 , R03A: Adrenergics, inhalants , 86 , 14.3 , R03AC: Selective beta-2 agonists , 43 , 7.2.

Techniques: Functional Assay, Blocking Assay, Analogues