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Biomedical subjects

R A Berg

Publications and source records attributed to R A Berg.

At least 109 records · Page 6Linked to original sources
PubMed1988

Material properties of living soft tissue composites.

Collagen sponges seeded with fibroblasts have been used as a soft tissue substitute in wound healing applications. This biomaterial is a good in vitro analog of a connective tissue. Therefore, analysis of the properties of this material may be useful for theoretically modeling soft tissues. Stress-strain curves for such cell-seeded collagen sponges were measured to determine composite stiffness and ultimate tensile strength. Theoretical modeling was done by defining a particle-reinforced matrix using the composite sphere model. A system of uniaxially oriented fibers was then introduced to this equivalent homogeneous media and material properties were determined using the composite cylinder model. Geometric averaging was performed to yield the stiffness and Poissons' ratio for a composite with randomly oriented fibers. Inputs to the model were constituent material properties, cell volume fraction, and fiber volume fraction. From theoretical results, material properties of soft tissues and their substitutes depend on fiber mechanical properties and volume fraction and not cellular mechanical properties and volume fraction. Therefore, the increase in experimentally observed composite stiffness with increased cell number was due to deposition of newly synthesized stiffer collagen fibers, and not due to the physical presence of cells themselves.

Collagen↗
PubMed1988

Behaviour of fibroblasts and epidermal cells cultivated on analogues of extracellular matrix.

A porous collagen sponge can be used for supporting epidermal cells and fibroblasts in order to manufacture an artificial skin. Fibroblasts were grown on analogues of extracellular matrix containing collagen and glycosaminoglycans and/or glycoproteins. Cell replication, and also infiltration of fibroblasts, were enhanced by the presence of hyaluronic acid and/or fibronectin. Epidermal cells grown on a collagen sponge have been characterized by microscopic observations. Epidermal cells on the surface of the sponge showed an incomplete differentiation in comparison to normal skin; clumps of epidermal cells were found in the interior of the sponge. Epidermal cell replication was enhanced in the presence of collagen sponge seeded with fibroblasts.

Animals↗
PubMed1988

Sexual abuse: somatic and emotional reactions.

A chart review and telephone interview of 72 sexual abuse victims was conducted to determine if children and adolescents who are victims of sexual abuse suffer from symptoms similar to the "rape trauma syndrome," which has been reported in adults. Symptoms similar to the rape trauma syndrome were found in 48 of the 72 abused children and only 26 of the matched control group, p less than .01. Common somatic complaints in the sexual abuse patients included dysuria, vaginal discharge and chronic abdominal pain. Some of the emotional and behavioral problems noted during the follow-up period among the sexual abuse patients included sleep problems, runaway behavior, and suicide attempts. The duration of abuse and age of the victim at the time of abuse significantly affected the frequency of reported somatic symptoms, but the type of abuse and type of assailant did not significantly affect the frequency of reported somatic and emotional reactions. There was no difference in the occurrence of school problems and early pregnancy between sexually abused patients and controls. Since 67% of all sexually abused patients suffered from emotional and somatic reactions, close follow-up of these patients is indicated.

Adaptation, Psychological↗
PubMed1988

Diagnostic parameters of an odd-even item short-form of the Luria-Nebraska Neuropsychological Battery.

The current study examined the concurrent validity of a new short form of Luria-Nebraska Neuropsychological Battery (LNNB). LNNB profiles of 100 subjects of a mixed brain damaged, psychiatric and normal pool were obtained from previously published sources. Levy corrected correlations among both the odd and even number short-forms with the full test ranged from.81 to.90. For the entire sample, absolute numbers of scales above the critical level, number of cases following within a given range of the critical level, and number of cases within each subscale exceeding the critical level were computed. Chi square values revealed significant differences only for the visual, reading, and memory subscales exceeding the critical level.

Journal Article↗
PubMed1988

Neutrophil response following intratracheal instillation of collagen peptides into rat lungs.

Inflammatory lung diseases are associated with destruction of interstitial collagen and release of degraded collagen fragments into the lower respiratory tract. To determine whether degraded collagen might be one factor mediating the cellular influx, we measured polymorphonuclear leukocytes (PMN) in bronchoalveolar lavage fluid following intratracheal instillation of collagen peptides. The responses to collagenous peptides derived from collagen digested with bacterial collagenase and to the collagenous-like polytripeptide (proline-proline-glycine) were examined. Both types of collagen peptides were chemotactic for human PMN in vitro. Two days following instillation of 5 mg collagenous peptides, we observed a threefold increase in the percentage of PMN in lavage fluid with a maximum (fivefold) response on day 6. Since other chemoattractants produce a response within hours when instilled intratracheally, we postulated that the late neutrophil influx produced with collagen may be the result of production of a neutrophil chemoattractant by alveolar macrophages. Alveolar macrophages treated with collagenous or collagenous-like peptides released PMN chemotactic factors, and the time course of release of chemotactic activity by alveolar macrophages in vitro correlated with the in vivo finding of a 2-6-day delay in PMN accumulation in the lungs. These observations are consistent with the idea that collagen peptides may stimulate alveolar macrophages to produce chemotactic factors for neutrophils. This mechanism may play a role in the accumulation of phagocytic cells in the lung following injury.

Animals↗
PubMed1988

Bone-cell-stimulating substance.

An acid-pepsin extract of ground cancellous calf bone contains a bone-cell-stimulating substance (BCSS) that is active in the intact rat and in bone cultures. A single injection of a suspension of BCSS adjacent to the radius-ulna complex of a rat significantly increased appositional bone formation in a dose-dependent manner. The pattern of new bone deposition differed from that caused by nonspecific irritants. A suspension or water-soluble extract of BCSS significantly increased DNA and collagen synthesis in organ cultures of diaphyseal pieces of 17-day-old fetal chick tibiae. Whether or not BCSS is different from any of the other bone-derived growth factors has not been established.

Animals↗
PubMed1988

Fibroblast and epidermal cell-type I collagen interactions: cell culture and human studies.

Fibroblast and epidermal cell-type I collagen sponge interactions were studied in cell culture as well as in humans. In cell culture, fibroblasts were observed to migrate and proliferate throughout a type I collagen sponge containing either hyaluronic acid (HA) or fibronectin (FN). Fibroblasts accumulated in the center of the pores in sponges containing HA and appeared to surround themselves with newly synthesized extracellular matrix. In sponges containing FN, fibroblasts attached to and elongated along the collagen fibers of the sponge. In the absence of FN or HA protein synthesis of fibroblasts appeared to be inhibited by the presence of the type I collagen sponge. Epidermal cells grown on plastic or on type I collagen, formed sheets. Epidermal cells grown on a collagen sponge morphologically appeared different than cells grown on plastic. The type I collagen matrix studied in cell culture was applied to dermal wounds of patients with pressure ulcers in order to evaluate its effect on dermal wound healing. The areas of ulcers treated for 6 weeks with a type I collagen sponge decreased by about 40% compared with no change in the areas of untreated controls. Preliminary results suggest that a type I collagen sponge is a biocompatible substrate with fibroblasts and epidermal cells and may be effective in enhancing healing of chronic skin ulcers.

Animals↗
PubMed1987

Fibroblast growth on a porous collagen sponge containing hyaluronic acid and fibronectin.

We have previously shown that the presence of fibronectin (FN) and/or hyaluronic acid (HA) in a 3-dimensional type I collagen sponge enhances wound healing in vivo. In the present study the same material was used as a support for growth of fibroblasts in vitro. Using radiochemical techniques, scanning electron and light microscopy, the properties of fibroblasts cultured on the collagen sponge or on the sponge containing HA or FN have been compared with cultures grown on plastic dishes. Fibroblast replication and collagen synthesis were higher on plastic than on the collagen sponge. In the presence of HA or FN the entire thickness of the sponge was infiltrated by fibroblasts which rapidly replicated. The presence of HA or FN increased synthesis of collagen which was largely deposited around cells.

Animals↗
PubMed1987

Damage and repair of lung connective tissue in rats exposed to toxic levels of oxygen.

We studied damage and repair of lung connective tissue in rats exposed to toxic amounts of oxygen by measuring lung content of collagen and elastin and the number of collagen fragments in lung lavage fluid after exposure to 98% O2 for 60 h. Lung collagen was decreased 17%, and collagen fragments in lavage fluid were increased 4- to 5-fold in O2-exposed rats compared with those in control rats. No biochemical evidence of elastin degradation was found. Mild emphysematous changes and a leftward shift of fluid-filled, pressure-volume curves were induced within 2 wk after recovery from exposure to O2. Administration of the lathyrogen beta-aminopropionitrile worsened the emphysematous lesion produced by hyperoxia, suggesting that replacement of connective tissue during repair limits the extent of emphysema. We conclude that lung collagen is degraded and an emphysematous lesion is produced by relatively short exposure to toxic amounts of oxygen.

Aging↗
PubMed1987

Degradation of collagen in lung tissue slices exposed to hyperoxia.

Exposure of animals to oxidant gases produces a mild emphysema, and O2-derived free radicals are capable of degrading connective tissues in vitro. It is postulated that degradation of connective tissue by O2-derived free radicals leads to emphysema in these models. To determine whether exposure of lung tissue slices to an oxidant gas results in degradation of collagen and to investigate factors mediating this degradation, we exposed lung tissue slices from normal rats to hyperoxia (95% O2, 5% CO2) and measured hydroxyproline release into the medium. After a 4-h exposure, the hydroxyproline released was 5.3 +/- 0.2 micrograms/g lung tissue (n = 10) in normoxia and 8.1 +/- 0.6 micrograms/g tissue (n = 13) in hyperoxia (p less than 0.05), suggesting degradation of collagen. The addition of 0.1% trypsin to the initial incubation medium caused a synergistic increase in hydroxyproline release from O2-exposed slices: normoxia/trypsin, 46.2 +/- 3.6 micrograms/g tissue (n = 10); hyperoxia/trypsin, 61.4 +/- 3.6 micrograms/g tissue (n = 11) (p less than 0.05). The addition of proteinase inhibitors completely suppressed the O2-induced release of hydroxyproline, suggesting that proteolytic enzymes are involved in hyperoxia-mediated degradation of lung collagen.

Animals↗
PubMed1986

Decreased synthesis and increased intracellular degradation of newly synthesized collagen in freshly isolated chick tendon cells incubated with monensin.

The synthesis and secretion of procollagen in embryonic chick tendon fibroblasts in suspension culture were inhibited with the carboxylic ionophore monensin. The synthesis of procollagen was inhibited by 50% in a 2-h exposure to 0.1 microM monensin and was inhibited by 70% in a 6-h exposure to 0.1 microM monensin. Secretion of procollagen was inhibited by greater than 90% in the 0.1 microM monensin-treated cultures and was totally inhibited by higher doses of the reagent. A cellular pool of collagenase-digestible peptides was demonstrated in the control cells, the level of which was elevated 3-4 times in the monensin-treated cultures. In order to determine whether the secretory and synthesis block caused by monensin inhibited intracellular degradation of newly synthesized collagen, the hydroxy[14C]proline in degraded collagen fragments present in control and monensin-treated cultures was determined and compared to the total hydroxy[14C]proline synthesized in each culture. The intracellular degradation of newly synthesized, pulse-labeled collagen was shown to proceed at rates comparable to those seen in the control cultures. The monensin-treated cells degraded pulse-labeled newly synthesized collagen nearly twice as long as the controls, resulting in an overall increase in the fraction of newly synthesized collagen that was degraded. These findings suggest that force generation in the activated cross-bridge cycle may occur as a result of an actin-attached cross-bridge transition between these two orientations.

Animals↗
PubMed1986

Regulation of collagen production and collagen mRNA amounts in fibroblasts in response to culture conditions.

Collagen synthesis and mRNA amounts for the alpha 1 and alpha 2 polypeptide chains of Type I collagen were measured in embryonic-chick tendons and in tendon cells both in suspension and in primary cultures. The percentage of protein production represented by collagen in suspension-cultured cells was initially the same as in the intact tendon; however, on an hourly basis, there was actually a steady decline in collagen production by suspended cells. Collagen production in primary cultures of chick tendon fibroblasts was decreased when compared with intact tendon, even though ascorbate-supplemented primary cultures were able to maintain higher rates of collagen production than were non-supplemented cultures. The amounts of mRNA for alpha 1(I) and alpha 2(I) polypeptide chains of collagen responded in similar fashions to different culture conditions and were compared with the amounts of mRNA for beta-actin. In primary cultures the available alpha 1 and alpha 2 collagen mRNAs support proportionately higher collagen production than in the intact tendon. However, the ratio of alpha 1/alpha 2 mRNA and polypeptide-chain synthesis did not remain 2:1, but increased with the concomitant production of Type I trimers composed of three alpha 1 chains. Removal of fibroblasts from their environment in vivo appears to alter the amounts of mRNA for alpha 1 and alpha 2 chains and to alter the utilization of those mRNAs for polypeptide synthesis.

Animals↗
PubMed1986

A novel isoform of cytoplasmic actin that binds poly-L-proline.

An actin-like protein was purified to apparent homogeneity from chick-embryo homogenates and chick-embryo fibroblasts by the use of poly-L-proline-agarose affinity chromatography; we therefore refer to this protein as PBP (poly-L-proline-binding protein). PBP binds to deoxyribonuclease-agarose, co-migrates with known actin standards on SDS/polyacrylamide-gel electrophoresis, and has an amino acid composition similar to that of actin. Linear peptide maps after digestion with Staphylococcus aureus proteinase reveal its apparent homology with gamma-actin; however, isoelectric-focusing experiments show that PBP is clearly more acidic than any of the three major isoforms of actin. PBP polymerizes in the presence of ATP to form fibrillar structures resembling actin paracrystalline aggregates. In chick-embryo fibroblasts, immunofluorescence with antibodies to PBP shows that its distribution is cytoplasmic: perinuclear staining of the cytoplasm, generalized cytoplasmic staining and peripheral fibrillar structures are evident. In contrast, antibodies specific for the (alpha, gamma)-actins reveal the typical stress fibre structures characteristic of fibroblastic cells. PBP appears to constitute a novel isoform of cellular actin, distinct from the known actin isoforms in terms of its lower isoelectric point, its ability to bind poly-L-proline and its distinct subcellular localization.

Actins↗
PubMed1986

Kinetics of intracellular degradation of newly synthesized collagen.

The objective of this work was to determine the time dependence of the basal component of intracellular degradation of newly synthesized collagen. Chick embryo tendon fibroblasts were incubated with [14C]proline, and degradation was quantified by measuring hydroxy[14C]proline in a low molecular weight fraction. When cultures were pulse labeled for 15 min and then incubated under chase conditions for 105 min, the amount of degraded collagen attained a value equal to approximately 20% of the amount synthesized during the labeling period; the data were fit with a simple exponential function that had a 40-min rise time and a 12-min lag time. In continuously labeled cultures, the rates of collagen synthesis and secretion reached constant values within 15 and 45 min, respectively. Degradation products were first detected 6-9 min after collagen synthesis began and were transported out of the cells more rapidly than intact collagenous molecules; however, percent degradation increased slowly and did not reach a constant value even after 240 min of incubation. Since collagen degradation lags collagen synthesis, it follows that degradation is a posttranslational, rather than a cotranslational, process, and since degradation and secretion are kinetically distinguishable, it follows that they occur in parallel pathways. A simple nonlinear model for posttranslational processing of collagen is proposed.

Animals↗
PubMed1986

Chemotactic activity of collagen-like polypeptides for human peripheral blood neutrophils.

Damage to interstitial connective tissue is associated with the rapid accumulation of monocytes and neutrophils at the site of injury. To study the role of collagen fragments in neutrophil migration, we analyzed the chemotactic properties of peptide fragments of bovine collagen digested with bacterial collagenase or cyanogen bromide and small molecular weight synthetic polypeptides containing proline (Pro), hydroxyproline (Hyp), and glycine (Gly), the major amino acids that comprise collagen. Using the Boyden chamber and under agarose techniques, we found that collagen fragments were as potent in inducing chemotaxis in neutrophils as the bacterial-derived peptide f-met-leu-phe. The synthetic polytripeptides (Pro-Pro-Gly)5 and (Pro-Hyp-Gly)5 were found to be equipotent in inducing chemotaxis, producing a maximal induction of chemotaxis at 5-10 nM. This suggests that Hyp, the unique imino acid found in collagen, is not required for chemotactic activity. Increasing the length of the synthetic tripeptide from 5 to 10 subunits decreased its chemotactic activity, while the single tripeptide subunit (Pro-Hyp-Gly)1 was the least active peptide, inducing a maximal response at 100 nM. To study the structural requirements for chemotaxis, Pro-Hyp-Gly tripeptides were synthesized with modifications at the N and C terminals ends. Addition of a methyl group to the carboxyl of Gly to form an ester enhanced the chemotactic activity of the peptide by 50%, while substitutions on the amino terminus with an acetyl group decreased the chemotactic activity by 50%. Substitution on the amino terminus with a Boc group decreased the chemotactic activity by 100%. These results indicate that there are specific structural requirements for chemotaxis induced by peptides having a collagen-like sequence of amino acids.

Chemotactic Factors↗
PubMed1986

Vasculitis in a suspected AIDS patient.

A 29-year-old male Haitian refugee had generalized lymphadenopathy, weight loss, and bilateral lung infiltrates diagnosed by transbronchial lung biopsy as tuberculosis. He had previously been labeled as having "pre-AIDS," which led to multiple suicide attempts. Four months later, cyanosis and gangrene of both lower extremities necessitated amputation, which revealed vasculitis.

Acquired Immunodeficiency Syndrome↗
PubMed1985

Characterization of the oligosaccharides of prolyl hydroxylase, a microsomal glycoprotein.

Prolyl hydroxylase is a tetrameric glycoprotein that catalyzes a vital posttranslational modification in the biosynthesis of collagen. The enzyme purified from whole chick embryos (WCE) possesses two nonidentical subunits, alpha and beta, and has been shown by several techniques to reside in the endoplasmic reticulum of chick embryo fibroblasts. The studies described here demonstrate that the larger of the two subunits (alpha) exists in two forms in chick embryo fibroblasts (CEF); these two forms differ in carbohydrate content. The larger alpha subunit, alpha', contains two N-linked high mannose oligosaccharides, each containing eight mannose units; the smaller subunit, alpha, contains a single seven-mannose N-linked oligosaccharide. Both oligosaccharides could be cleaved by endo-beta-N-acetylglucosaminidase H and completely digested with alpha-mannosidase to yield mannosyl-N-acetylglucosamine.

Animals↗