Management of acute anemia in a Jehovah's Witness patient with acute lymphoblastic leukemia with polymerized bovine hemoglobin-based oxygen carrier: a case report and review of literature.

Background: Management of severe symptomatic anemia is exceptionally challenging when blood transfusions cannot be administered. Use of hemoglobin (Hb)-based oxygen carriers as "bridging treatment" can be an option.

Case Report: This is a report of a 36-year-old Jehovah's Witness diagnosed with acute lymphoblastic leukemia (ALL) who developed severe symptomatic anemia (Hb as low as 3.

HBOC-201 vasoactivity in a phase III clinical trial in orthopedic surgery subjects--extrapolation of potential risk for acute trauma trials.

Background: Vasoactivity has hampered progress of hemoglobin-based oxygen carriers (HBOCs) due to concern for adverse blood pressure responses and secondary complications. A recent formulation, highly polymerized HBOC-201 (Biopure, Cambridge, MA), has been found to be less vasoactive than prior less polymerized formulations, and to improve outcome in animal models of hemorrhagic shock (HS) compared with standard resuscitation fluids. HBOCs are envisioned to have life- saving potential for severe trauma patients for whom death due to HS is common despite transport to level I trauma centers.

Basic science focus on blood substitutes: a summary of the NHLBI Division of Blood Diseases and Resources Working Group Workshop, March 1, 2006.

In March 2006, a workshop sponsored by the National Heart, Lung, and Blood Institute was convened to identify the role of basic science research in clarifying issues that are impeding progress in the development of hemoglobin-based oxygen carrying (HBOC) solutions. These discussions resulted in a consensus that, although HBOCs have shown clinical promise, various side effects have inhibited further development and regulatory approval, with cardiovascular events being of particular concern. As a consequence, workshop participants focused on formulating research recommendations to better understand and mitigate these side effects.

Lycium barbarum glycoconjugates: effect on human skin and cultured dermal fibroblasts.

Lycium barbarum L. (Solanaceae) glycoconjugates (LbGp) display an interesting array of anti-apoptotic and antioxidant properties, which may be beneficial for human skin. We therefore set out to determine the effects of LbGp in full-thickness human skin, and in dermal fibroblasts.

The epithelial mucin, MUC1, is expressed on resting T lymphocytes and can function as a negative regulator of T cell activation.

MUC1 is a mucinous glycoprotein which is normally expressed on the surface of a variety of epithelia and aberrantly overexpressed on some human tumors. In this report, we demonstrate that the epithelial mucin, MUC1, is expressed on resting human peripheral blood T cells and two leukemia T cell lines, Jurkat and Hut 78. Crosslinking of MUC1 on peripheral blood T cells by plate-bound anti-MUC1 (DF3-P) antibody inhibits cell proliferation, IL-2 and GM-CSF production, and up-regulation of the IL-2 receptor upon anti-CD3 stimulation.

Persistent transgene expression in mouse liver following in vivo gene transfer with a delta E1/delta E4 adenovirus vector.

Extensive in vivo gene transfer studies in animal models and human gene therapy clinical trials with E1-deleted adenovirus vectors have demonstrated transience of transgene expression due to direct cytopathic effects of the vectors and host immune response to virally expressed proteins. In order to overcome these difficulties, we have recently developed packaging cell lines which support the growth of adenovirus vectors containing lethal deletions in both E1 and E4 gene regions. Here we demonstrate that use of E1/E4-deleted adenovirus vectors leads to prolonged in vivo transgene expression due to elimination of cytopathic effects and significant reduction of virus-specific immune response.

cDNA-derived primary structure of the 25-kDa subunit of canine microsomal signal peptidase complex.

We purified the 25-kDa subunit of the pentameric signal peptidase complex (SPC 25) from canine pancreas microsomes and utilized amino acid sequence data from tryptic fragments for its molecular cloning and sequencing. Its primary structure (calculated molecular mass of 25,831 Da) shows the presence of two hydrophobic domains of which one is likely to serve as a transmembrane segment. Consistent with being the only SPC subunit with at least one intrachain disulfide bond, SPC 25 contains 4 Cys residues.

Gene targeting in normal somatic cells: inactivation of the interferon-gamma receptor in myoblasts.

Gene targeting in somatic cells represents a potentially powerful method for gene therapy, yet with the exception of pluripotent mouse embryonic stem (ES) cells, homologous recombination has not been reported for a well characterized, non-transformed mammalian cell. Applying a highly efficient strategy for targeting an integral membrane protein--the interferon gamma receptor--in ES cells, we have used homologous recombination to target a non-transformed somatic cell, the mouse myoblast, and to compare targeting efficiencies in these two cell types. Gene-targeted myoblasts display the properties of normal cells including normal morphology, ability to differentiate in vitro, stable diploid karyotype, inability to form colonies in soft agar and lack of tumorigenicity in nude mice.

A subunit of mammalian signal peptidase is homologous to yeast SEC11 protein.

Canine signal peptidase consists of a complex of five proteins (Evans, A. E., Gilmore, R.

Do motilin and pancreatic polypeptide regulate duodenal bile acid delivery?

The plasma levels of the enteric hormones, motilin and pancreatic polypeptide, cycle in association with fasting intestinal motility and are altered by feeding. Intravenous administration of motilin causes gallbladder contraction and increased sphincter of Oddi phasic motor activity, whereas pancreatic polypeptide causes gallbladder relaxation. To determine if endogenous plasma levels of motilin and pancreatic polypeptide control sphincter of Oddi and gallbladder motility, and regulate duodenal bile acid delivery, we measured during fasting and after feeding the correlation between (a) changes in plasma motilin or pancreatic polypeptide, and (b) the duodenal delivery of a steady-state hepatic output of radiolabelled bile acid.

Cytoskeleton and thyroglobulin expression change during transformation of thyroid epithelium to mesenchyme-like cells.

In considering the mechanism of transformation of epithelium to mesenchyme in the embryo, it is generally assumed that the ability to give rise to fibroblast-like cells is lost as epithelia mature. We reported previously that a definitive embryonic epithelium, that of the anterior lens, gives rise to freely migrating mesenchyme-like cells when suspended in type I collagen matrices. Here, we show that a highly differentiated epithelium that expresses cytokeratin changes to a vimentin cytoskeleton and loses thyroglobulin during epithelial-mesenchymal transformation induced by suspension in collagen gel.

Cytodifferentiation and tissue phenotype change during transformation of embryonic lens epithelium to mesenchyme-like cells in vitro.

A number of adult and embryonic epithelia, when suspended within native type I collagen gels, give rise to elongate bipolar cells that migrate freely within the three-dimensional matrix. The morphology of these newly formed mesenchyme-like cells is indistinguishable from "true" mesenchymal cells at the light and ultrastructural level. In this report, we extend previous observations on the transformation of embryonic avian lens epithelium to mesenchyme-like cells.

Epithelia suspended in collagen gels can lose polarity and express characteristics of migrating mesenchymal cells.

This study of epithelial-mesenchymal transformation and epithelial cell polarity in vitro reveals that environmental conditions can have a profound effect on the epithelial phenotype, cell shape, and polarity as expressed by the presence of apical and basal surfaces. A number of different adult and embryonic epithelia were suspended within native collagen gels. Under these conditions, cells elongate, detach from the explants, and migrate as individual cells within the three-dimensional lattice, a previously unknown property of well-differentiated epithelia.

Synthesis and distribution of cytoskeletal elements in endothelial cells as a function of cell growth and organization.

Confluent cultures of adult bovine aortic endothelial (ABAE), corneal endothelial (BCE), and fetal bovine heart endothelial (FBHE) cells form a monolayer of highly flattened, closely apposed, and nonoverlapping cells. In ABAE and BCE cultures, this is associated with a 50-fold decrease in the rate of DNA sythesis and correlates with a 14-fold decrease in protein synthesis. In contrast, in confluent FBHE cultures only partial decreases in the rates of DNA synthesis (6-fold) and protein synthesis (3-fold) are observed.

Corneal endothelial replacement. I. In vitro formation of an endothelial monolayer.

Cultured bovine corneal endothelial cells were seeded onto a corneal button denuded of its own endothelium. After an incubation time of 30 min to 1 hr, these cultured cells were able to repopulate and reconstitute a new endothelium over Descemet's membrane. Three basic cellular processes were involved in the formation of the new endothelial layer.

The production and localization of laminin in cultured vascular and corneal endothelial cells.

The production and localization of laminin, as a function of cell density (sparse versus confluent cultures) and growth stage (actively growing versus resting cultures), has been compared on the cell surfaces of cultured vascular and corneal endothelial cells. Comparison of the abilities of the two types of cells to secrete laminin and fibronectin into their incubation medium reveals that vascular endothelial cells can secrete 20-fold as much laminin as can corneal endothelial cells. In contrast, both cell types produce comparable amounts of fibronectin.

Vascular endothelial cells maintained in the absence of fibroblast growth factor undergo structural and functional alterations that are incompatible with their in vivo differentiated properties.

Vascular endothelial cells cultured in the presence of fibroblast growth factor (FGF) adopt at confluence a morphological appearance similar to that of the vascular endothelium in vivo. Similarly, their apical cell surface is, as in vivo, nonthrombogenic. In contrast, when the cultures are maintained in the absence of FGF, the cells undergo within two to three passages structural and functional alterations that are incompatible with their in vivo morphological appearance and physiological function.

Transplantation of cultured bovine corneal endothelial cells to species with nonregenerative endothelium. The cat as an experimental model.

The in vivo transplantation of cultured bovine corneal endothelial cells has been attempted in the cat. Cat corneas denuded of their endothelium were coated with bovine corneal endothelial cells previously maintained in tissue culture. When grafted back into cat recipients, the corneal buttons remained clear with no edema.

Transplantation of cultured bovine corneal endothelial cells to rabbit cornea: clinical implications for human studies.

Rabbit corneas denuded of their endothelium were coated with bovine corneal endothelial cells (from steers) previously maintained in tissue culture for short (20 generations) or prolonged (200 generations) periods. When grafted back into female rabbits, the corneal buttons remained clear and showed no edema. In contrast, denuded corneas coated with bovine keratocytes and grafted into rabbits became opaque and edematous within 7 days and remained so thereafter.