Long-term outcome of transanal irrigation for children with spina bifida.

STUDY DESIGN: Experimental, prospective study. OBJECTIVES: We evaluated the long-term clinical efficacy of transanal irrigation (TAI) and its effect on the quality of life of spina bifida children and their caregivers. SETTING: Republic of Korea. METHOD: Forty-four spina bifida pediatric patients with constipation, fecal incontinence or both, underwent a TAI program at our spina bifida clinic between December 2010 and October 2013. The children and their caregivers were evaluated using a self-administered questionnaire before TAI and at 3 months and 3 years after initiation of the program. RESULTS: Successful treatment outcome was achieved in 38 (86.4%) children after a mean follow-up duration of 33 months (range, 30-36). The mean number of fecal incontinence episodes per week, the number of diaper changes and the total time for bowel care per day before the program decreased at the latest follow-up examination from 7.3 to 0.4 (P<0.001), 1.6 to 0.2 (P<0.001) and 29.2 to 19.4 min (P=0.038), respectively. These results remained constant from short-term follow-up at 3 months to 3 years. Caregivers and children could go out more often (P=0.002), and the emotional impact of bowel care on caregivers decreased (P<0.001). The reported mean overall satisfaction with TAI was 8/10. The common adverse effect during TAI was abdominal discomfort (60.5%). CONCLUSION: We observed a sustained significant improvement in defecation symptoms and quality of life for 3 years in spina bifida children who underwent continuous TAI.

PR-39 coordinates changes in vascular smooth muscle cell adhesive strength and locomotion by modulating cell surface heparan sulfate-matrix interactions.

PR-39 is proline-rich peptide produced at sites of tissue injury. While the functional properties of this peptide have not been fully defined, PR-39 may be an important regulator of processes related to cell-matrix adhesion since it reportedly upregulates syndecan-4, which is a critical determinant of focal adhesion formation. The ability of PR-39 to modulate the adhesion and chemokinetic migration behavior of arterial smooth muscle cells (SMCs) in a fashion coordinated with syndecan-4 expression was investigated. Treatment of SMCs with PR-39 did not alter syndecan-1 mRNA, but did induce a two-fold increase in syndecan-4 mRNA (P < 0.0001) and significantly enhanced cell surface expression of both syndecan-4 (P < 0.01) and heparan sulfate (HS) (P < 0.05). These observations were consistent with an observed increase in cell-matrix adhesive strength (P < 0.05) and a reduction in cell speed (P < 0.01) on fibronectin-coated substrates. Incubation of PR-39 treated cells with a soluble fibronectin derived heparin-binding peptide, as a competitive inhibitor of heparan sulfate/matrix interactions, abolished these effects. These data suggest that PR-39 mediated alterations of cell adhesion and motility may be related, in part, to the increased expression of heparan sulfate glycosaminoglycans (GAGs) that accompany the upregulation of cell surface syndecan-4. Furthermore, this investigation supports the notion that factors which control syndecan-4 expression may play an important role in regulating adhesion related cell processes.

Culture results and amputation rates in high-pressure paint gun injuries of the hand.

High-pressure paint gun injuries have been well described in the literature, and the use of antibiotics is recommended as part of their management. However, there is no scientific evidence to support the use of antibiotics. In addition, the type of paint injected (water- versus oil-based) has never been investigated to determine the extent of morbidity resulting from these injuries. This study examines the organisms cultured in wounds resulting from these injuries and whether the type of paint injected had an influence on amputation rates. Charts of 35 patients with high-pressure paint gun injuries to their hands were reviewed. The amputation rate was 50% with oil-based paints and 0% with water-based paints. Forty-seven percent of wound cultures were positive, with gram-negative bacteria found in 58% of isolates. Our findings support the use of antibiotics, which should cover both gram-positive and gram-negative organisms.

Soluble heparin-binding peptides regulate chemokinesis and cell adhesive forces.

The ability of a soluble heparin-binding peptide sequence derived from fibronectin to modulate the adhesion and chemokinetic migration behavior of arterial smooth muscle cells was assessed using a novel glass microsphere centrifugation assay and automated time-lapse fluorescence videomicroscopy, respectively. Treatment of cells grown on fibronectin-coated substrates with the soluble heparin-binding peptide resulted in the disassembly of focal adhesions, as assessed by immunohistochemical staining. These observations were consistent with an observed dose-dependent two- to fivefold reduction in cell-substrate adhesive strength (P < 0.001) and a biphasic effect on migration speed (P < 0.05). Moreover, heparin-binding peptides induced a twofold reduction (P < 0.01) in two-dimensional cell dispersion in the presence of a non-heparin-binding growth factor, platelet-derived growth factor-AB (PDGF-AB). Heparin-binding peptides were unable to mediate these effects when cells were grown on substrates lacking a heparin-binding domain. These data support the notion that competitive interactions between cell surface heparan sulfates with heparin-binding peptides may modulate chemokinetic cell migration behavior and other adhesion-related processes.

Cytomimetic biomaterials. 3. Preparation and transport studies of an alginate/amphiphilic copolymer/polymerized phospholipid film.

The significance of molecular design methodologies based upon membrane-mimetic systems lies in the ability to engineer robust materials of varying geometry with a high degree of reproducibility and molecular control over surface order and chemistry. However, non-covalently associated assemblies, in and of themselves, are often insufficiently robust for many applications. We have previously reported the in situ polymerization of a single phospholipid monolayer on a self-assembled film of octadecyltrichrolosilane (OTS) on glass, as well as the polymerization of phospholipids on an amphiphilic, dialkyl-containing terpolymer bound to a gold-coated silicon wafer. We now report the polymerization of a phospholipid monolayer assembly onto an alkylated hydrogel substrate with significant alteration in both surface chemistry and mass transport properties at the hydrogel-water interface. A general platform is thereby created for enhancing the control of either the local delivery of specific macromolecules or the immunoisolation barrier for many cell based therapies.

Alpha4beta1 and alpha5beta1 control cell migration on fibronectin by differentially regulating cell speed and motile cell phenotype.

Computer-aided time lapse fluorescence videomicroscopy was used to study single cell migration behavior of human aortic endothelial cells on fibronectin coated substrates of varying protein surface density. The role of receptors alpha5beta1, alpha(v)beta3, and alpha4beta1 in mediating cell adhesion and migration on fibronectin was characterized using integrin specific monoclonal antibodies. Matrix density had a direct effect on controlling the proportion of migrating cells and the directional persistence of cell movement (p<0.01). While there was relatively little influence of fibronectin surface density on absolute migration speed, the ability of endothelial cells to disperse over a surface, as measured by the dispersion coefficient, was biphasic with respect to the surface density of this matrix protein (p<0.005). Both cell speed and the proportion of migrating cells was controlled by alpha4beta1 (p<0.01). However, alpha5beta1 selectively regulated the transformation of stationary cells to those exhibiting motile behavior (p<0.05). Migratory responses on fibronectin were not influenced by blockade of the alpha(v)beta3 receptor. It is noteworthy that cell surface adhesive receptors which control commitment to a motile phenotype are not necessarily the same as those that control migration speed.

Characterization of single-cell migration using a computer-aided fluorescence time-lapse videomicroscopy system.

Single-cell assays of cell migration, while yielding dynamic measurements of cell position and morphology, are predominantly limited by the time required for data collection and analysis. Computer-aided fluorescence time-lapse videomicroscopy (CAFTiV) was developed in order to facilitate the tracking and rapid examination of large numbers of motile cells. The system combines time-lapse videomicroscopy with epifluorescence capability, which allows full automation of image capture, sorting, and analysis due to the low background in the fluorescence images. Utilizing the CAFTiV system, data analysis time was reduced from over 125 h to less than 1 labor minute. In addition, fluorescence imaging permits cell tracking in small-volume chambers (<100 microL), which is useful should the addition of expensive reagents be required. It is anticipated that the ability to characterize both biochemical and biophysical properties responsible for cell movement will be enhanced by this methodology.

Role of fibronectin and sulfated proteoglycans in endothelial cell migration on a cultured smooth muscle layer.

Endothelial cell (EC) migration is essential for the healing of denudation injuries to the vessel wall. This event occurs in vivo in a pericellular environment and should be regulated in part by juxtacrine and paracrine interactions with underlying smooth muscle cells (SMCs). To investigate the EC migration behavior over SMCs under direct cell-cell contact conditions, we have utilized human umbilical vein endothelial cells (HUVECs) grown to confluence on microcarrier beads and radiolabeled with chromium-51. The EC carrying beads were spread over a confluent human aortic SMC monolayer and cocultured for 48 hr, allowing the EC to migrate from beads onto the underlying SMC monolayer in the presence of sodium chlorate, cycloheximide (CHM), or anti-fibronectin monoclonal antibody (anti-FN mAb). The level of EC migration was quantified by counting the amount of radioactivity on the SMC layer after the removal of the beads. The presence of a SMC layer enhanced EC migration more than threefold (P < 0.05). Furthermore, EC migration was inhibited from 30 to 80% (P < 0.05) by sodium chlorate, CHM, and anti-FN mAb in a dose-dependent fashion. This model has shown that smooth muscle cells augment endothelial motility. Both fibronectin and sulfated proteoglycans released by ECs and SMCs likely play an important role in the regulation of EC migration behavior over SMCs. The method described of using radiolabeled ECs on microcarrier beads should prove to be a useful tool in the study of cell migration in a heterotypic cellular environment.