Biomedical subjects

M W Siegel

Publications and source records attributed to M W Siegel.

12 recordsLinked to original sources

PubMed1996

Distribution of DNA and alginate in purulent cystic fibrosis sputum: implications to pulmonary targeting strategies.

Cystic fibrosis (CF) patients frequently experience recurring airway infections characterized by thick, viscous sputum. The consistency and nature of these purulent secretions may produce a significant barrier to the successful delivery of drugs and gene therapy vectors designed to treat CF. We have carried out a series of in vitro studies to determine the distribution of two macromolecular components typically present in purulent sputum, bacterial alginate and neutrophil-derived DNA. Sputum samples were obtained from hospitalized CF patients. DNA and alginate were disrupted, respectively, by the in vitro additions of human recombinant deoxyribonuclease I (rhDNase) or alginate lyase prepared from a mucoid strain of Pseudomonas aeruginosa. N-acetyl-L-cysteine (acetylcysteine) was similarly used to collapse the mucin matrix of these samples for comparison. Using a centrifugation-based rheological method known as the compaction assay, a greater maximal response was observed for rhDNase compared to alginate lyase treatment. A simultaneous addition of these enzymes to purulent sputum produced an additive compaction response. Electron microscopy was used to identify alginate and DNA components within the mucin matrix of sputa and to evaluate changes following treatment with high concentrations of alginate lyase or rhDNase. DNA was more widely distributed throughout purulent samples than alginate. Differences in the distribution of DNA and alginate may explain, at least in part, the larger compaction response to rhDNase versus alginate lyase treatment. An improved understanding of DNA and alginate distribution within purulent CF sputum may lead to improvements in drug and vector delivery to airway epithelial cells.

Alginates↗

PubMed1995

Disruption of retinal axon ingrowth by ablation of embryonic mouse optic chiasm neurons.

Mouse retinal ganglion cell axons growing from the eye encounter embryonic neurons at the future site of the optic chiasm. After in vivo ablation of these chiasm neurons with a monoclonal antibody and complement, retinal axons did not cross the midline and stalled at approximately the entry site into the chiasm region. Thus, in the mouse, the presence of early-generated neurons that reside at the site of the future chiasm is required for formation of the optic chiasm by retinal ganglion cell axons.

Animals↗

PubMed1995

Frameless patient registration using ultrasonic imaging: a preliminary study.

Locating the patient accurately in real time is necessary for many medical robotics applications. Current solutions are expensive or inconvenient. We show progress toward solving these problems by using ultrasound to track computed tomography (CT) reference features in realtime. We compute the locations of target features in the ultrasonic images relative to the CT reference features.

Algorithms↗

PubMed1993

One systemic administration of transforming growth factor-beta 1 reverses age- or glucocorticoid-impaired wound healing.

The role of intravenously administered recombinant human transforming growth factor-beta 1 (rhTGF-beta 1) on the healing of incisional wounds in rats with impaired healing due to age or glucocorticoid administration was investigated. The administration of methylprednisolone to young adult rats decreased wound breaking strength to 50% of normal control. Breaking strength of incisional wounds from 19-mo-old rats was decreased approximately 27% compared with wounds from normal healing young adult rats. A single intravenous administration of rhTGF-beta 1 (100 or 500 micrograms/kg) increased wound breaking strength from old rats or young adult rats with glucocorticoid-induced impaired healing to levels similar to normal healing control animals when determined 7 d after injury. Even though the circulating half-life of systemically administered rhTGF-beta 1 is < 5 min, a sustained stimulatory effect on extracellular matrix secretion was evident in glucocorticoid-impaired rats when rhTGF-beta 1 was administered at the time of wounding, 4 h after wounding, or even 24 h before wounding. These observations indicate a previously unrecognized potential for the active form of TGF-beta 1 to profoundly influence the wound healing cascade after brief systemic exposure.

Aging↗

PubMed1992

Endocytosis of glycophospholipid-anchored and transmembrane forms of CD4 by different endocytic pathways.

A glycophosphatidylinositol (GPI)-anchored form of the CD4 receptor was constructed by fusing the extracellular domain of CD4 to the COOH-terminus of decay accelerating factor (DAF), containing a signal for GPI-anchor attachment. The internalization of GPI-linked CD4 (CD4DAF) was compared to that of transmembrane CD4 using both [125I]gp120 and anti-CD4 antibodies. We show that transmembrane CD4 is rapidly endocytosed in transfected CHO cells, while CD4DAF is internalized at a rate approximately 3-fold slower. Immunoelectron microscopy suggests that whereas transmembrane CD4 is endocytosed via clathrin-coated vesicles, CD4DAF enters cells by an alternative pathway involving non-coated microinvaginations of the plasma membrane. Following internalization CD4DAF recycles through a primaquine-insensitive compartment, whereas the recycling of transmembrane CD4 is inhibited by primaquine, suggesting that the two receptors may recycle from distinct populations of early endosomes. Colocalization of both CD4DAF and CD4 with an antibody against a lysosomal membrane protein suggests that the two endocytic pathways may converge.

Animals↗

PubMed1990

Male-specific beta-cell dysfunction and diabetes resulting from increased expression of a syngeneic MHC class I protein in the pancreata of transgenic mice.

It is well established that insulin-dependent diabetes (IDDM) is an autoimmune disease with a strong genetic link to the HLA locus. It is less well understood, however, how the destruction of the insulin-producing beta cells is effected and why neighboring non-beta islet cells are spared. Also incompletely explained are the observations that, unlike other autoimmune diseases such as multiple sclerosis, IDDM does not preferentially affect females, the incidence of the disease is highest among young adults, and there are temporal correlations between the onset of the disease and emotional trauma. We have addressed some of these questions by using transgenic mice that constitutively express the MHC class I antigen Dd in the beta cells of the pancreas. Although both male and female Ins.Dd mice expressed equivalent amounts of the Dd protein only the males developed diabetes. The diabetes in the males could be reversed by castration, and the normoglycemic females became diabetic following either ovariectomy and the implantation of a slow-release pellet containing testosterone or the inclusion of dexamethasone in the drinking water. In contrast, transgenic mice that expressed the herpes simplex virus type 1 glycoprotein D in the pancreatic beta cells were normoglycemic and showed no obvious histopathological consequences. The observation that the beta-cell dysfunction by the increased expression of the MHC class I protein Dd cannot be induced by the herpes viral protein suggests that the cellular damage is related to a specific structure or function of the MHC proteins.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1981

Post-embedding immunoperoxidase staining of glial fibrillary acidic protein for light and electron microscopy.

Post-embedding peroxidase-antiperoxidase methods to stain glial fibrillary acidic (GFA) protein in Araldite-embedded sections for light and electron microscopy were developed. The Jimpy mouse spinal cord was used because it is gliotic and contains abundant glial filaments and GFA protein. For light microscopy, specific staining was obtained in thick and in ultrathin sections mounted on glass following removal of the plastic with sodium ethoxide. Consistent specific staining for GFA protein in ultrathin sections mounted on nickel grids required partial removal of the plastic with 1% sodium ethoxide and further treatment with 2% sodium dodecyl sulfate (SDS).

Animals↗

PubMed1969