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M A Riederer-Henderson

Publications and source records attributed to M A Riederer-Henderson.

12 recordsLinked to original sources

Ultrasonic assessment of skin and surgical wounds utilizing backscatter acoustic techniques to estimate attenuation.

Backscatter acoustic techniques at high ultrasonic frequencies (10-40 MHz) were utilized to investigate the ultrasonic attenuation coefficient of normal skin and the relationship between attenuation and the healing of surgical wounds ranging in age from 9 to 49 days. The attenuation coefficient was calculated with measurements from depths of 0.5 and 1.0 mm, primarily in the reticular dermis. The values for control skin ranged from 6.0 Np cm-1 at 10 mHz to 19.6 Np cm-1 at 40 MHz and a corresponding slope of 0.45 Np cm-1 MHz-1. Wound attenuation initially increased with wound age from approximately 15% of control at day 9 to 30% of control at day 34 but did not continue to increase through day 49. During the same period, however, total collagen (as a percentage of wet weight) increased at a constant rate from approximately 45% of control at day 9 to 70% of control at day 49. Thus the attenuation coefficient in healing wounds over the time period studied may be sensitive to more than total collagen content in the tissue. It may be affected by other competing factors as wounds mature such as intermolecular cross-linking, collagen fiber bundle size, and structural arrangement of fiber bundles or other tissue constituents such as proteoglycans and elastin. The observations that wound integrated attenuation using backscatter techniques over the range of 10-40 MHz substantially increased in early wounds, did not increase beyond day 34, and remained significantly lower than control values over the entire healing period studied are in agreement with the results of an independent study of the same tissue using transmission techniques at 100 MHz.

Animals↗

Correlation of tissue constituents with the acoustic properties of skin and wound.

The purpose of this study was to compare measurements of ultrasound properties of skin and wound tissue with measurements of material properties such as total collagen concentration, acetic acid soluble collagen concentration, water concentration, and morphologic properties. Using a scanning laser acoustic microscope (SLAM), both ultrasonic speed and attenuation coefficient values were obtained for control skin (2-3 cm from the wound), for skin immediately adjacent to wounds (within 0.3 mm), as well as for wound tissue itself. The attenuation coefficient and speed measurements were lowest for wound tissue followed by adjacent skin and then control skin. As the wounds healed there appeared to be an increase in both speed and attenuation coefficient although the wound age at which these increases started and the length of time for which they continued varied from one dog to the next. The precision of duplicate sample measurement of wave speed was +/- 1.7% for control skin, whereas that for attenuation coefficient it was +/- 16%. Both ultrasonic speed and attenuation coefficient were directly correlated with tissue collagen concentration and inversely correlated with tissue water concentration (p less than 0.001). Attenuation coefficient correlated best (r = 0.73) with acetic acid soluble collagen concentration which reflects the changes in collagen taking place during the repair process. These attenuation measurements made at 100 MHz using the SLAM were compared for control skin and wound samples with measurements made at 10-40 MHz using backscatter acoustic techniques (BAT). The tissue samples analyzed by each ultrasound technique were from adjacent locations on the animals.

Acoustics↗

Neutral metalloprotease from tendons.

Tendon repair following trauma, rupture, or surgery involves both synthesis and degradation of collagen in order to reweave new collagen bundles in with the old. Using an in situ assay on polyacrylamide gels containing gelatin, we have identified protease activity from tendon tissue and from tendon cells in culture. A population of synovial cells from the epitenon surrounding the tendon as well as the tendon fibroblasts themselves were examined. The cells and the conditioned medium from both cell populations exhibited a major band of gelatin-degrading activity at 70 kdaltons and a minor band of activity at 60 kdaltons. When preparations were reacted with p-aminophenylmercuric acetate (APMA) before electrophoresis, a third band appeared at 63 kdaltons. The main band at 70 kdaltons comigrated with a [35S]methionine-radiolabeled protein band. Inhibitor and pH studies identified the enzymes as neutral metalloproteases requiring disulfide bonds for activity. No proteolytic activity was detected on casein-containing gels, ruling out the presence of stromelysin. Since electrophoresis in the presence of SDS would separate the metalloprotease from the smaller molecular weight inhibitor (TIMP), these in situ assays provide a sensitive screening system for gelatin-degrading enzymes present in tendon without prior removal of TIMP.

Animals↗

Effects of theophylline on expression of the long cilia phenotype in sand dollar blastulae.

Previously, increases in ciliary length have only been obtained through genetic mutation in Chlamydomonas or by incubation of swimming echinoderm blastulae in trypsin or elastase. We have found that the phenotypic switch from short to long cilia on sand dollar blastulae can also be effected by incubation in theophylline. Cilia detached from control blastulae have a mean length of 21 +/- 7 microns with 10% of the cilia being greater than 30 microns. Upon incubation in 10 mM theophylline additional long cilia appeared after 10 hours and by 24-32 hours 1/2-3/4 of the embryo was covered with long cilia. The percentage of long cilia increased to 65% with a mean length of 40.0 +/- 17.6 microns. Incubation in other methylxanthines, such as aminophylline, caffeine, or isobutylmethylxanthine, inhibited development but had no effect on ciliary length distribution. Dibutyryl cAMP, 8-bromoadenosine, and calcium ionophore also had no effect on ciliary length. Cyclic AMP levels were measured and showed only slight differences among controls and embryos incubated in trypsin, caffeine, or theophylline. These data suggest that theophylline may be altering ciliary length control through some mechanism other than elevations in cAMP.

Animals↗

Ultrasonic assessment of skin and wounds with the scanning laser acoustic microscope.

The aim of the present study was to test the hypothesis that ultrasonic propagation properties in skin and wound tissue would correlate with material properties such as collagen content, water content, and tensile strength of those tissues. Both ultrasonic speed and ultrasonic attenuation coefficient were directly correlated with tissue collagen content, [r = 0.80 and r = 0.56, respectively (p less than 0.001)]. In addition, ultrasonic speed and attenuation coefficient were inversely correlated with tissue water content, [r = -0.57 and r = -0.73, respectively (p less than 0.001)]. Tensile strength also correlated very significantly with ultrasonic speed (r = 0.90, p less than 0.001), and significantly with attenuation coefficient (r = 0.58, p less than 0.001). The results demonstrate the feasibility of using ultrasound for noninvasively determining the material properties of biologic tissues including healing cutaneous wounds.

Animals↗

Phenotypic switching to long cilia effected by various proteases: results with Dendraster excentricus and Stronglyocentrotus purpuratus blastulae.

Incubation in trypsin effects a phenotypic switch from short to long cilia (greater than 30 micron) in hatched blastulae of the sea urchin, Arbacia punctulata. To determine how trypsin causes such a switch we tested whether the phenomenon was unique to the species, Arbacia, and to the protease, trypsin. With two other echinoderm species, the sand dollar, Dendraster excentricus, and the sea urchin, Strongylocentrotus purpuratus, trypsin incubation increased the percentage of long cilia. During incubation of D. excentricus in trypsin, the percentage of long cilia increased progressively from the normal 10% long cilia of the apical tuft to 45-50% long cilia covering 1/2-3/4 of the embryo. With S. purpuratus blastulae, however, the percentage of long cilia was lower (32-40%) and the results were more variable. Of the additional proteases tested with D. excentricus, elastase was more effective than trypsin in terms of the percentage of long cilia (58%), the mean length, and the broad distribution of lengths formed. Thermolysin was about as effective as trypsin but chymotrypsin was much less so. Thus, increases in ciliary length were not unique to a particular echinoderm species or to incubation in trypsin. The magnitude of the change in length distribution, however, was species- and enzyme-dependent. An extracellular or membrane component with differential susceptibility to various proteases may, therefore, be involved in altering ciliary length.

Animals↗

A low-serum medium for tendon cells: effects of growth factors on tendon cell growth and collagen production.

Primary avian tendon cells maintain a higher percentage of net collagen synthesis when cultured in low serum concentrations than in high. However, under these conditions the cells grow slowly and can only be subcultured one or two times. We therefore examined various growth factors for their effects on tendon cell multiplication in order to develop a new medium for these cells. Of the growth factors tested, epidermal growth factor, insulin, transferrin, and selenium each stimulated tendon cell division. We also investigated how these factors affected collagen synthesis, and found that they all increased both collagen and noncollagen protein synthesis equally, thus leaving the percentage of protein synthesis devoted to collagen the same. When combined with 0.2% fetal bovine serum in Ham's F12 medium, epidermal growth factor and insulin together stimulated cell multiplication to a level comparable to that of cells grown in Ham's F12 plus 10% serum. Cells could also be successfully subcultured in this medium. Thus, by using selected growth factors we have reduced significantly the serum requirements of cultured tendon cells without affecting population doubling time or subculture capability. This low-serum medium should prove useful for the study of the regulation of collagen biosynthesis.

Animals↗

Attachment and extracellular matrix differences between tendon and synovial fibroblastic cells.

Fibroblasts of the synovium of sheathed tendons were isolated, and their biochemical properties were compared with those of the fibroblasts of the remaining tendon. The synovial cells had a lower attachment efficiency than did the tendon cells. On the day of cell isolation the synovial cells synthesized collagen as 10% of their total protein, whereas the tendon cells synthesized 30% collagen. After growth in fetal bovine serum (FBS), the percentage of collagen synthesized by both populations decreased; however, the synovial cells still made less collagen than did the tendon cells (5 versus 11%). On the basis of cyanogen bromide peptide analysis, the synovial cells were found to synthesize Types I and III collagen in primary culture, whereas the tendon cells synthesized only Type I. The synovial cells also synthesized two to three times less sulfated glycosaminoglycans in culture than did the tendon cells. Thus, the two cell populations differed in attachment efficiency and in their biosynthesis of collagen and sulfated glycosaminoglycans. These differences reflect extracellular matrix differences that have been observed in the tendon in vivo. In addition, the results augment existing data showing that not all fibroblasts have identical phenotypes.

Animals↗

H(2) metabolism in photosynthetic organisms: I. Dark h(2) evolution and uptake by algae and mosses.

Dark H(2) metabolism was studied in marine and fresh water red algae, the green alga, Chlamydomonas, and mosses. A time variable and temperature-sensitive anaerobic incubation was required prior to H(2) evolution. H(2) evolution was sensitive to disalicylidenepropanediamine. An immediate H(2) uptake was observed in these algae. Immediate dark H(2) uptake but no evolution was observed in the mosses.A cell-free hydrogenase preparation was obtained from anaerobically adapted Chlamydomonas reinhardii by means of sonic oscillation. The hydrogenase was not sedimented at 100,000g. It catalyzed the reduction of methylene blue, p-benzoquinone, NAD, NADP, but not spinach ferredoxin. H(2) evolution was noted with dithionite and with reduced methyl viologen as donors but not with reduced spinach ferredoxin. Similarly, hydrogenase activities were not affected by disalicylidenepropanediamine. The pH optima for H(2) evolution and for H(2) uptake were 7.2 and 7.5 to 9.5, respectively. Extracts prepared from the anaerobically adapted red alga, Chondrus crispus, and the moss, Leptobryum pyriforme, consumed but did not evolve H(2). Uptake was slightly stimulated by methylene blue. It is proposed that red algae and mosses appear to metabolize H(2) by a different pathway than Chlamydomonas.

Journal Article↗