Communications in construction design

Construction design has become an increasingly complex synthesis activity for which effective solutions depend upon co-operative participation by a number of people. Thus communication, including the integration of specialised knowledge and negotiation of differences between team members, is a vital process for collaborative design. A questionnaire survey was initially conducted to investigate communication issues and problems, which had been highlighted from a review of the literature, in current construction design. The results confirmed that communication among the different construction team members is often difficult although of paramount important to design outcomes. Based on these results, case studies have been carried out to gain further insights into communication issues and problems, and explore why and how they are caused. Through the application of multiple approaches, a model has been developed, which suggests strategies that may help participants communicate more effectively and ultimately improve the quality of construction design outcomes

Cell-Specific Cre Strains For Genetic Manipulation in Salivary Glands

<div><p>The secretory acinar cells of the salivary gland are essential for saliva secretion, but are also the cell type preferentially lost following radiation treatment for head and neck cancer. The source of replacement acinar cells is currently a matter of debate. There is evidence for the presence of adult stem cells located within specific ductal regions of the salivary glands, but our laboratory recently demonstrated that differentiated acinar cells are maintained without significant stem cell contribution. To enable further investigation of salivary gland cell lineages and their origins, we generated three cell-specific Cre driver mouse strains. For genetic manipulation in acinar cells, an inducible Cre recombinase (Cre-ER) was targeted to the prolactin-induced protein (<i>Pip</i>) gene locus. Targeting of the <i>Dcpp1</i> gene, encoding demilune cell and parotid protein, labels intercalated duct cells, a putative site of salivary gland stem cells, and serous demilune cells of the sublingual gland. Duct cell-specific Cre expression was attempted by targeting the inducible Cre to the <i>Tcfcp2l1</i> gene locus. Using the R26^{Tomato Red} reporter mouse, we demonstrate that these strains direct inducible, cell-specific expression. Genetic tracing of acinar cells using Pip^GCE supports the recent finding that differentiated acinar cells clonally expand. Moreover, tracing of intercalated duct cells expressing Dcpp^GCE confirms evidence of duct cell proliferation, but further analysis is required to establish that renewal of secretory acinar cells is dependent on stem cells within these ducts.</p></div

Characterization of <i>Pip</i>^<i>GCE</i> knock-in allele.

<p><b>(A)</b> Generation of <i>Pip</i>^<i>GCE</i> knock-in mice. <i>Pip</i> genomic structure and restriction map is shown at the top. White box represents the non-coding exon sequences and filled boxes, the coding sequences. Thick bars show the sequences used to generate the homologous arms in the targeting vector. Gray box represents 3’ external probe used for Southern blotting. Arrows indicate positions of genotyping PCR primers (An3’ and PipR). <b>(B-E)</b> Analysis of Cre expression in mice after 3 days of tamoxifen treatment, followed by a 3-day chase. <b>(B)</b> Frozen sections were prepared from submandibular gland (SMG); activation of Cre results in expression of Tomato red reporter (TdT) (red); Scale bar = 50 <i>μ</i>m. No Cre activity is detected in <b>(C)</b> parotid (Par), <b>(D)</b> lacrimal gland (Lac) or <b>(E)</b> sublingual gland (SLG). Nuclei are stained with DAPI (blue). Scale bars = 25<i>μ</i>m. <b>(F)</b> Section from <i>Pip</i>^<i>GCE/+</i><i>;R26</i>^<i>TdT/+</i> SMG at 3 days after tamoxifen treatment. Single labeled acinar cells (red) co-localize with antibody to Nkcc1 (green). Scale bar = 50<i>μ</i>m <b>(G)</b> Section from <i>Pip</i>^<i>GCE/+</i><i>;R26</i>^<i>TdT/+</i> SMG at P9, isolated 3 days after tamoxifen administration. Positively labeled acinar cells are red. Nuclei are stained with DAPI. Scale bar = 25<i>μ</i>m <b>(H)</b> Section from <i>Pip</i>^<i>GCE/+</i><i>;R26</i>^<i>TdT/+</i> SMG at 3 months after tamoxifen treatment, co-stained with antibody to Nkcc1 (green) to label acinar cells. Labeled acinar cells have expanded to clones (red). Scale bar = 50<i>μ</i>m <b>(I)</b> Section from <i>Pip</i>^<i>GCE/+</i><i>;R26</i>^<i>TdT/+</i> SMG after 3 month chase shows expansion of labeled acinar cells into clones (arrowheads). <i>3d</i>, 3 days chase; <i>3mos</i>, 3 month chase; <i>d</i>, duct; Scale bar = 50 <i>μ</i>m.</p

Schematic diagram of general salivary gland structure.

<p>Secretory acinar cells are arranged in clusters, known as acini, which produce primary saliva. The smallest intercalated ducts conduct saliva from the acini to the striated, and excretory ducts. Sites of inducible Cre drivers are indicated, color-coded for each strain. <i>Dcpp1</i>, gene encoding demilune cell and parotid protein; <i>Pip</i>, gene encoding prolactin-inducible protein; <i>Tcf</i>, gene encoding Tcfcp2l1 transcription factor.</p

Characterization of <i>Dcpp1</i>^<i>GCE</i> knock-in allele.

<p><b>(A)</b> Generation of <i>Dcpp1</i>^<i>GCE</i> knock-in mice. <i>Dcpp1</i> genomic structure and restriction map is shown at the top. White box represents the non-coding exon sequences and filled boxes, the coding sequences. Thick bars show the sequences used to generate the homologous arms in the targeting vector. Arrows indicate positions of external long-range PCR primers (5’ external primer and GCE primer) and internal primers (An3’ and Dcpp1R) used for genotyping. <b>(B)</b> Analysis of Cre expression in sublingual gland (SLG) of <i>Dcpp1</i>^<i>GCE/+</i><i>;R26</i>^<i>TdT/+</i> mice after 3 days of tamoxifen treatment, followed by a 3-day chase. Activation of Cre results in expression of Tomato red reporter (TdT) (red). <b>(C)</b> Higher magnification of labeled SLG cells reveals the morphology of serous demilunes (arrowheads). <b>(D)</b> Antibody to Nkcc1 labels SLG acinar cell membranes, and co-localizes with TdT-labeled serous demilune cell (yellow; arrowhead). <b>(E)</b> Analysis of Cre expression in parotid gland (Par) of <i>Dcpp1</i>^<i>GCE/+</i><i>;R26</i>^<i>TdT/+</i> mice after 3 days of tamoxifen treatment, followed by a 3-day chase. Activation of Cre results in expression of TdT (red) in small clusters of intercalated duct cells (arrowheads). <b>(F)</b> Higher magnification of TdT-labeled (red) intercalated duct cells (arrowhead). Nuclei are stained with DAPI (blue). <b>(G)</b> Antibody to Nkcc1 labels acinar cells (green). TdT-positive cells (red) do not co-localize with acinar cells, but are found within the smallest intercalated ducts (arrowheads). <b>(H)</b> Section from <i>Dcpp1</i>^<i>GCE/+</i><i>;R26</i>^<i>TdT/+</i> parotid gland after 3 days of tamoxifen treatment, followed by a 3-month chase. TdT-positive cells (red) are clustered in duct-like structures (arrows). <b>(I)</b> At 3 months chase, TdT-labeled cells (red) derived from Dcpp1-expressing cells are clustered in intercalated ducts (arrows). Some Dcpp1-labeled cells may overlap with acinar cells labeled with antibody to Nkcc1 (green; arrowhead). Nuclei are stained with DAPI (blue). <i>3d</i>, 3 days chase; <i>3mos</i>, 3 month chase; Scale bars = 50<i>μ</i>m (B,E,H); = 25<i>μ</i>m (C,D,F,G); = 20<i>μ</i>m (I).</p

Additional file 1: of A synthetic cell-penetrating peptide derived from nuclear localization signal of EPS8 exerts anticancer activity against acute myeloid leukemia

Changes in EGF/PDGF signaling pathway targets analyzed with a RT2 profiler™ PCR assay. A Array layout of the RT2 profiler™ PCR assay. B Changes in EGF/PDGF signaling pathway targets in KG1α/sh1 cells compared with those in KG1α/NC cells. (TIFF 6966 kb

Generation of <i>Pou4f1</i> and <i>Pou4f2</i> conditional knockout alleles.

<p>(A) Targeting strategy for generating <i>Pou4f1</i> conditional null allele. (B) Targeting strategy for generating <i>Pou4f2</i> conditional null allele. (C) Southern blot confirmed selection of <i>Pou4f1^cko</i> allele and PCR genotyping of <i>Pou4f1^loxP</i> heterozygous and homozygous. (D) Southern blot confirmed selection of <i>Pou4f2^cko</i> allele and PCR genotyping of <i>Pou4f2^loxP</i> heterozygous and homozygous. White boxes are the non-coding exon sequences and black boxes are the coding sequences. Thick bars are the sequences used to generate the homologous arms in the targeting vector. Restriction enzyme sites: A, <i>AvrII</i>; B, <i>BamHI</i>; E, <i>EcoRI</i>; H, <i>HindIII</i>; N, <i>NotI</i>; S, <i>SacII</i>; X, <i>XbaI</i>.</p

No significant change in the number of RGCs in adult <i>Pou4f2CKO</i> mice.

<p>Retina flat mounts of control and <i>Pou4f2CKO</i> mice were collected after tamoxifen injection. (A–C) Two weeks after tamoxifen treatment, RGCs labeled by TUJ1 (A) and ISL1 (B) and DAPI in GCL (C); (D–F) Four weeks after tamoxifen treatment, RGCs labeled by TUJ1 (D) and ISL1 (E) and DAPI in GCL (F); (G–I) Three months after tamoxifen treatment, RGCs labeled by TUJ1 (G) and ISL1 (H) and DAPI in GCL (I); (J–L) Six months after tamoxifen treatment, RGCs labeled by TUJ1 (J) and ISL1 (K) and DAPI in GCL (L); (M) Quantification results of each cell marker in 1,600 µm². Scale bar equals to 100 µm.</p

No significant change in optic nerve diameter, axonal elongation and glia activation in control and doubleCKO mice.

<p>Both control and doubleCKO mice were collected six months after tamoxifen treatment. No significant change in the optic nerve diameter (A and D), SMI32 immunolabeling (B and E) and GFAP immunolabeling (C and F) is seen between control and doubleCKO mice. Scale bar equals to 100 µm.</p

Cell counts of caspase3⁺ cells after CONC in <i>DoubleCKO</i> and control retinas.

<p>Cell counts of caspase3⁺ cells after CONC in <i>DoubleCKO</i> and control retinas.</p